CN104663723B - Prevent and treat complex micro organism fungicide of downy mildew of fruits and vegetables and preparation method thereof - Google Patents
Prevent and treat complex micro organism fungicide of downy mildew of fruits and vegetables and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of complex micro organism fungicide for preventing and treating downy mildew of fruits and vegetables, containing two kinds of effective bacterial components, it is respectively Bacillus cercus Bacillus cereus BCJB01 and bacillus megaterium Bacillus megaterium BMJBN02, downy mildew of garpe can be prevented and treated, cucumber downy mildew can be prevented and treated again, the composite bacteria agent is the suspending agent compounded using two kinds of different biocontrol bacterial strains of the mode of action, to grape and cucumber downy mildew more than 88.76%.
Description
Technical field
The present invention relates to biological technical field, more particularly to a kind of complex micro organism fungicide for preventing and treating downy mildew of fruits and vegetables and
Its preparation method.
Background technology
Downy mildew is serious to the harm of Curcurbitaceae melon crop.The pathogenic bacteria of downy mildew of cucurbits are Pseudoperonospora cubensis
[Pseudoperonosporacubensis (Berk.et Curt.) Rostov], it is Cucumis and muskmelon to infect frequency highest
Platymiscium, more than 80 cultivated cucumber of countries and regions muskmelon of a countries and regions more than 50 is fallen ill extensively.The pathogen exists
Under natural conditions, muskmelon and cucumber are not only infected, also to watermelon cucurbitaceous, pumpkin, sponge gourd, wax gourd, cucurbit and serpent melon etc. 12
Kind of melon crop causes harm (willow swallow, Xu Yongyang etc..Muskmelon downy mildew progress [J], Chinese melon dish, 2011,24 (3)：
38-43)。
Downy mildew of garpe is by Plasmopara viticola [Plasmoparaviticola (Berk.et Curtis.) Berl.et
De Toni.] (Chen Hao, Hu Liang are refined etc. for a kind of worldwide obligatory parasitism disease that causes.Bacillus subtillis B-FS01 is to grape
The prevention effect [J] of downy mildew.Plant protection, 2011,37 (6)：194-197).There is generation China viny region, is harm
One of grape leaf diseases of most serious, serious economic loss is caused to grape production.
From Bordeaux mixture invention in 1882 so far, chemical prevention and control be always control downy mildew harm, it is popular, cause disaster it is most main
The measure wanted, most fruits and vegetables producing regions need repeatedly, a large amount of could success using chemical agent and causes disaster at its harm of prevention and control, and this is simultaneously
Also increase the risk (Qin Wentao that chemical agent produces side effect.Plasmopara viticola rapid molecular detection method studies [D],
The Chinese Academy of Agricultural Sciences).Therefore, exploitation is found safer to people and animals, to ecological environment is without destructiveness and can make up chemical agriculture
The pharmaceutical preparations of some defects of medicine are current problem demanding prompt solutions.
In recent years, country encourages and advocates control of plant disease and to develop to environmental protection direction.Therefore, downy mildew is biological
The exploitation of agricultural chemicals has realistic meaning.At present, there is the extract solution class of the biological reagent including plant of good preventive effect, micro- to downy mildew
Biological and its endotoxin, Compost Extracts class, plant growth regulator and bio-fertilizer class, bioprotein class and some wettable powders
Agent class.
The content of the invention
It is big that the present invention is directed to grape and cucumber downy mildew difficulty of prevention and cure in solving agricultural production currently with biological control, into
This height, chemical pesticide residual it is exceeded the problems such as and provide it is a kind of prevent and treat downy mildew of fruits and vegetables complex micro organism fungicide and its preparation side
Method, can prevent and treat downy mildew of garpe, and cucumber downy mildew can be prevented and treated again, and the composite bacteria agent is using two kinds of different lifes of the mode of action
The suspending agent of anti-bacterial strain compounding, to grape and cucumber downy mildew more than 88.76%.
Bacillus cercus Bacillus cereus BCJB01 of the present invention are white by Liaocheng of Shandong Province Gaotang County
Separating obtained in field of vegetables, culture presevation number is CGMCC NO.10477, with the unique effect for decomposing downy mildew zoospore.
Bacillus megaterium Bacillus megaterium BMJBN02 in the Chinese chive field soil of Shandong Province Shouguang by separating institute
, culture presevation number is CGMCC NO.10478, with the effect for deforming downy mildew sporangium.
Complex micro organism fungicide and preparation method thereof the technical scheme of preventing and treating downy mildew of fruits and vegetables of the invention is, the preventing and treating
The complex micro organism fungicide of downy mildew of fruits and vegetables, is respectively Bacillus cercus Bacillus containing two kinds of effective bacterial components
Cereus BCJB01, in being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on January 30th, 2015
The heart, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：100101, deposit number is CGMCC NO.10477, bacterium
The Latin title planted is Bacillus cereus, joins the microorganism (strain) of evidence：BCJB01.
Bacillus megaterium Bacillus megaterium BMJBN02, Chinese micro- life is preserved on January 30th, 2015
Thing culture presevation administration committee common micro-organisms center, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：
100101, deposit number is CGMCC NO.10478, and the Latin title of strain is Bacillus megaterium, ginseng evidence
Microorganism (strain)：BMJBN02.
When the complex micro organism fungicide prevents and treats downy mildew of garpe, Bacillus cercus Bacillus cereus BCJB01 and
The number of viable ratio of bacillus megaterium Bacillus megaterium BMJBN02 is 1~3:2；During preventing and treating cucumber downy mildew,
Bacillus cercus Bacillus cereus BCJB01's and bacillus megaterium Bacillus megaterium BMJBN02
Number of viable ratio is 1:0.5~2.
Preferably, when the complex micro organism fungicide prevents and treats downy mildew of garpe, Bacillus cercus Bacillus cereus
The number of viable ratio of BCJB01 and bacillus megaterium Bacillus megaterium BMJBN02 is 3:2；Preventing and treating cucumber downy mildew
When sick, Bacillus cercus Bacillus cereus BCJB01 and bacillus megaterium Bacillus megaterium
The number of viable ratio of BMJBN02 is 1:2.
The preparation method of the complex micro organism fungicide of described preventing and treating downy mildew of fruits and vegetables, comprises the following steps：
(1) culture of seed liquor：By Bacillus cercus Bacillus cereus BCJB01 and bacillus megaterium
Bacillus megaterium BMJBN02 are inoculated into LB liquid medium respectively, and 30 DEG C, 200rpm shaken cultivations 12~18 are small
When, the OD values at 600nm are measured by sampling, returned to zero with nonvaccinated LB liquid medium in continuous mode；
(2) preparation of zymotic fluid：
1. the preparation of Bacillus cercus Bacillus cereus BCJB01 zymotic fluids：When the OD values in step (1) are
When 0.5~0.8, seed liquor is pressed 1:100 volume ratios add the culture tank equipped with fermentation medium；
2. the preparation of bacillus megaterium Bacillus megaterium BMJBN02 zymotic fluids：OD in step (1)
Be worth for 0.5~0.8 when, by seed liquor press 1:150 volume ratios add the culture tank equipped with fermentation medium；
(3) the mixing compounding of complex microorganism：Bacillus cercus Bacillus cereus BCJB01 fermentations are taken respectively
Liquid and bacillus megaterium Bacillus megaterium BMJBN02 fermentating liquid volumes compare 1:1 or 1:3 carry out mixing compounding；
(4) preparation of complex microorganism suspending agent：Step (3) is mixed into mixing compositional liquor and the surface-active that compounding is obtained
Agent, preservative, suspension aids and thickener are mixed with suspending agent.
LB liquid medium formula in step (1)：1%NaCl, 1% peptone, 0.5% dusty yeast, pH7.0~7.5.
The preparation of the 1. Bacillus cercus Bacillus cereus BCJB01 zymotic fluids in step (2), condition of culture
For：30 DEG C of cultivation temperature, rotating speed 200r/min, fermentation time is 24~36h, and zymotic fluid pH maintains 7.0, dissolved oxygen amount 20%,
5~7m of throughput3/ hour, 0.05~0.1KPa of tank pressure, spore production rate is more than 90%, count plate 8 × 109CFU·mL-1；Hair
Ferment culture medium prescription：Corn flour 2%, bean cake powder 1%, dusty yeast 0.3%, CaCO30.3%, MgSO4·7H2O 0.02%,
K2HPO4·3H2O 0.2%, KH2PO40.2%, pH7.0~7.2.
The preparation of the 2. bacillus megaterium Bacillus megaterium BMJBN02 zymotic fluids in step (2), culture
Condition is：30 DEG C of cultivation temperature, rotating speed 220r/min, fermentation time is 36~48h, and zymotic fluid PH maintains 7.0, dissolved oxygen amount
20%, 5~7m of throughput3/ hour, 0.05~0.1KPa of tank pressure, spore production rate is more than 80%, count plate 5.33 ×
109CFU·mL-1；Fermentative medium formula：Corn flour 2%, bean cake powder 1%, dusty yeast 0.3%, CaCl20.1%, MgSO4·
7H2O 0.05%, K2HPO4·3H2O 0.3%, KH2PO40.2%, pH7.0~7.5.
Suspension aids are ammonium sulfate in step (4), and preservative is glycine, and surfactant is organosilicon BD-3077, are increased
Thick dose is xanthans.
In Bacillus cercus Bacillus cereus BCJB01 zymotic fluids and bacillus megaterium in step (4)
In Bacillus megaterium BMJBN02 zymotic fluid mixing compositional liquors, by mass percentage add ammonium sulfate 6%~
8%, glycine 2%~5%, BD-30770.05%~0.1%, xanthans 0.2%~0.3% is obtained, living in gained suspending agent
Bacterium is counted as 6.0~8.0 × 109CFU·mL-1；The consumption of above each component is radix to mix the quality of compositional liquor.
The preparation of step (4) complex microorganism suspending agent is specially：First ammonium sulfate stirring is descended in addition mixing compositional liquor,
After after ammonium sulfate all dissolving, preservative glycine, surfactant organosilicon BD-3077, thickening are sequentially added under agitation
Agent xanthans, stirs.
Complex micro organism fungicide of preventing and treating downy mildew of fruits and vegetables of the invention and preparation method thereof has the beneficial effect that：The present invention is
Specifically designed for the biological prevention and control agent of harm downy mildew of fruits and vegetables, due to being biological agent, it is completely absent because chemicals is using bringing
A series of problems, be thus advantageous to the No-harmful apple orchard of fruits and vegetables, not only for user saves spending, and improve
The quality of fruits and vegetables production, is conducive to exporting extra earning.
Greenhouse experiments show：Complex micro organism fungicide of the invention can significantly reduce the generation of downy mildew of garpe.Field
Experiment shows, when complex micro organism fungicide of the invention dilutes 100 times of liquid, the prevention effect to downy mildew of garpe is 89.37%.
Under greenhouse experiment, the protection prevention effect to cucumber downy mildew is 88.76%.
Surfactant organosilicon BD-3077 of the invention, the main chain of BD-3077 is very submissive in actual applications, its
Intermolecular force ratio hydrocarbon wants much weaker, i.e., compared with the hydrocarbon with molecular weight, its viscosity is low, surface
Tension force is weak, and surface can be small, and film forming ability is strong.Therefore, the low surface tension and low-surface-energy of BD-3077 for its bring it is hydrophobic, disappear
The items excellent properties such as bubble, foam stabilization, anti-sticking, lubrication, glazing；Meanwhile, BD-3077 has also had surfactant and synergy concurrently
The effect of agent.
Specific embodiment：
For a better understanding of the present invention, technical scheme is described in detail with instantiation below, but originally
Invention is not limited thereto.
Embodiment 1
Strain isolation and identification
1st, Bacillus cercus Bacillus cereus BCJB01 and BMJBN02 of the present invention, are chatted by Shandong Province
Chinese cabbage Tanaka is separating obtained for city Gaotang County, and culture presevation number is CGMCC NO.10477, with decomposition downy mildew zoospore
Unique effect
Bacillus megaterium Bacillus megaterium BMJBN02 in the Chinese chive field soil of Shandong Province Shouguang by separating institute
, culture presevation number is CGMCC NO.10478, with the effect for deforming downy mildew sporangium.
2nd, identification of strains
Determined using physiological and biochemical property and method that 16SrDNA sequence alignment analysis are combined by bacterial strain BCJB01 and
BMJBN02 is identified respectively as Bacillus cercus (Bacillus cereus) and bacillus megaterium (Bacillus
megaterium).Meanwhile, the single bacterium colony that will isolate and purify at the same carry out V.P experiments, mycoderm formed experiment, Nagler's reaction,
Seven identifications of key character such as Starch Hydrolysis experiment, casein hydrolysis test, aesculin, cellobiose, determine foundation and join
Write according to eastern show pearl, Cai Miaoying etc.《Common bacteria system identification handbook》.
Embodiment 2
It is prepared by complex microorganism suspending agent
The preparation of 1 seed liquor：By above-mentioned biocontrol microorganisms Bacillus cercus Bacillus cereus BCJB01 and huge bud
Spore bacillus Bacillus megaterium BMJBN02 are inoculated into LB liquid medium, 30 DEG C, 220rpm shaken cultivations 12 respectively
~18 hours, the OD values at 600nm are measured by sampling in superclean bench, are adjusted with nonvaccinated LB liquid medium in continuous mode
Zero；
The preparation of 2 zymotic fluids：
1. the preparation of Bacillus cercus Bacillus cereus BCJB01 zymotic fluids：When nutrient solution OD values be 0.5~
When 0.8, seed liquor is pressed 1:100 volume ratios add the culture tank equipped with fermentation medium.30 DEG C of cultivation temperature, rotating speed 200r/
Min, fermentation time is 24~36h, and zymotic fluid pH maintains 7.0, dissolved oxygen amount 20%, 5~7m of throughput3/ hour, tank pressure 0.05
~0.1KPa, spore production rate is more than 90%, count plate 8 × 109CFU·mL-1；Fermentative medium formula：Corn flour 2%, beans
Dregs of rice powder 1%, dusty yeast 0.3%, CaCO30.3%, MgSO4·7H2O 0.02%, K2HPO4·3H2O 0.2%,
KH2PO40.2%, pH7.0~7.2；
2. the preparation of bacillus megaterium Bacillus megaterium BMJBN02 zymotic fluids：OD in step (1)
Be worth for 0.5~0.8 when, by seed liquor press 1:150 volume ratios add the culture tank equipped with fermentation medium；30 DEG C of cultivation temperature,
Rotating speed 220r/min, fermentation time is 36~48h, and zymotic fluid PH maintains 7.0, dissolved oxygen amount 20%, 5~7m of throughput3/ small
When, 0.05~0.1KPa of tank pressure, spore production rate is more than 80%, count plate 5.33 × 109CFU·mL-1；Fermented and cultured basigamy
Side：Corn flour 2%, bean cake powder 1%, dusty yeast 0.3%, CaCl20.1%, MgSO4·7H2O 0.05%, K2HPO4·3H2O
0.3%, KH2PO40.2%, pH7.0~7.5.
The mixing compounding of 3 complex microorganisms：BCJB01 and BMJBN02 fermentating liquid volumes are taken respectively compares 1：1、1：2、1：3 enter
Row mixing compounding, then, mixing compositional liquor is determined to Plasmopara viticola and the inhibitory action of cucumber downy mildew by leaf disk method,
The optimal mixing compound proportion of screening.
Specific testing program is as follows：
Experiment sets 5 treatment, 100 times of dilutions of zymotic fluid of biocontrol bacterial strain BCJB01,100 times of the zymotic fluid of BMJBN02
Dilution, BCJB01 and BMJBN02 fermentating liquid volumes compare 1：1、1：2、1：3 100 times of dilutions of mixing compositional liquor, sterilized water pair
According to.
Collection incidence of leaf and fresh non-incidence of leaf (grape variety is damp mountain one, and cucumber variety is imperial jade seal), hair
Sick blade brushes off the mould layer on scab under flowing water with writing brush, and 22 DEG C, relative humidity 90%, each 12h alternate cultures of brightness are waited to grow
When going out new mould layer, with the lower germ sporangium of distillation washing, suspension is made into, concentration is controlled to 1 × 105CFU·mL-1, 4 DEG C
Save backup.
Beaten between the vein of non-incidence of leaf with diameter 1.5cm card punch and take leaf dish, be respectively placed in and contain different disposal
1min is soaked in liquid, then blade back faces up on the blotting paper for being placed in soaked with immersion leaf dish same concentrations liquid respectively,
The μ L of sporangia suspension 20 that will be prepared are inoculated in leaf dish center, are subsequently placed in illumination box, 22 DEG C, relative humidity
90%, each 12h alternate cultures of brightness.Incidence is investigated after inoculation 7d.
Severity graded index：0 grade, disease-free spot；1 grade, lesion area accounts for leaf dish area below 5%；3 grades, scab
Area accounts for leaf dish area 5%-20%；5 grades, lesion area accounts for leaf dish area 20%-50%；7 grades, lesion area accounts for leaf dish area
More than 50%.
Disease index=∑ (morbidity leaf dish number × relative extreme at different levels)/(total leaf dish number × 7 of investigation) × 100
Inhibiting rate (%)=[(control disease refers to-process that disease refers to)/control disease refers to] × 100
The mixing compositional liquor of table 1 most preferably mixes the screening test of compound proportion
As shown in Table 1, BCJB01 and BMJBN02 fermentating liquid volumes ratio is 1：1 (thalline quantity ratio be 3：2) when, to grape
Preferably, inhibiting rate is 96.38% to the inhibition of downy mildew；BCJB01 and BMJBN02 fermentating liquid volumes ratio is 1：3 (thalline numbers
Amount is than being 1：2) when, the inhibition to cucumber downy mildew is best, and inhibiting rate is 90.53%.
The preparation of 4 complex microorganism suspending agents：Compositional liquor and surfactant, preservative, suspension will most preferably be mixed respectively
Auxiliary agent and thickener etc. are mixed with suspending agent, wherein prevent and treat downy mildew of garpe is complex microorganism suspending agent I, prevent and treat cucumber
Downy mildew is complex microorganism suspending agent II.
Suspension aids of the present invention are ammonium sulfate, and preservative is glycine, and surfactant is organosilicon BD-3077, thickening
Agent is xanthans.
Complex microorganism suspending agent of the invention, in BCJB01 and BMJBN02 most preferably mix compositional liquor, by quality percentage
Than adding ammonium sulfate 6%~8%, glycine 2%~5%, BD-30770.05%~0.1%, xanthans 0.2%~0.3% to make
, count plate is 6.0~8.0 × 10 in gained suspending agent9CFU·mL-1.The consumption of above each component is mixing compositional liquor
Quality be radix.
Embodiment 3
Preventing and treating of the complex microorganism suspending agent to downy mildew
Field plot trial of the 1 complex microorganism suspending agent I to downy mildew of garpe
Experiment is carried out in the Bio-Centers vineyard of Shandong Scientific Research Academy, and kind is damp mountain one.
Experiment is by the field test criterion design of medicine inspecting institute of the Ministry of Agriculture, totally 4 treatment：Complex microorganism suspending agent I uses clear water
100 and 600 times of liquid, 20% 600 times of Propamocarb aqua liquid, clear water control are diluted to respectively.Using spray-on process dispenser, in the afternoon
16：Carried out after 00, often process 10 plants, 3 repetitions, RANDOMIZED BLOCK DESIGN.Pumping is newly climing then for every cell random searching 10, from
Above 10 leaves of lower investigation, record the sick numbers of sheets at different levels.The same leaf disk method of blade grade scale.Start to apply when there is fragmentary morbidity in field
Medicine, interval medication in 7 days 1 time, continuous use 3 times.The disease index after 5 days investigation medicines after last time dispenser.Put down during experiment
Equal 28 DEG C of temperature, precipitation 210mm.
Disease index=∑ (the sick level number of sheets × this grade of typical value)/investigation total number of sheets × morbidity is highest
Preventive effect (%)=[1- (and before the medicine of check plot disease refer to × treatment region medicine after disease refer to)/(disease refers to × processes after the medicine of check plot
Disease refers to before area's medicine)] × 100
Field efficacy of the complex microorganism suspending agent I of table 2 to downy mildew of garpe
As shown in Table 2, complex microorganism suspending agent I has good field control effect to downy mildew of garpe, in dilution 100
During times liquid, preventive effect is 89.37%；Preventive effect is 76.88% during 600 times of liquid of dilution, better than the Propamocarb aqua of comparison medicament 20%
Preventive effect.
Greenhouse controlling experiment of the 2 complex microorganism suspending agents II to cucumber downy mildew
Experiment is carried out in the Bio-Centers greenhouse of Shandong Scientific Research Academy, and kind is imperial jade seal.Two kinds of not same-action effects are pressed in experiment
Fruit is carried out, respectively protecting effect experiment and therapeutic effect experiment.
Protecting effect is tested：When cucumber seedling just appears true leaf, complex microorganism suspending agent II is sprayed the 1st time.The 1st
Sprayed for 2nd time when true leaf is considerable.The concentration of complex microorganism suspending agent II is respectively 100 and 600 times of liquid, if 20% Propamocarb aqua
600 times of liquid and clear water are control.Bacterium of downy mildew of cucumber sporangia suspension is inoculated with after 10d, disease index and guarantor are recorded after morbidity
Shield effect.
Therapeutic effect is tested：Cucumber downy mildew sporangia suspension is carried out into complete stool spraying to plant with sprayer at dusk to connect
Kind, with preservative film moisturizing 24h, medicament is sprayed at the initial stage of a disease, sprayed again every 6 days 1 time, spray 3 times altogether, record disease index and treatment
Effect.
Disease index=∑ (the sick level number of sheets × this grade of typical value)/investigation total number of sheets × morbidity superlative degree prevention effect=
[(the sick level index of control-treatment disease level index) the sick level index of/control] × 100% data statistic analysis：Statistical analysis is used
EXCEL2003 and SPSS19.0 softwares are carried out, and Multiple range test is using the new multipole difference methods of inspection of Deng Kenshi.
Greenhouse controlling experiment of the complex microorganism suspending agent II of table 3 to cucumber downy mildew
As shown in Table 3, complex microorganism suspending agent II has good prevention effect to cucumber downy mildew, in 100 times of dilution
During liquid, protecting effect preventive effect is 88.76%；Therapeutic effect preventive effect is 71.30%.Protecting effect preventive effect is during 600 times of liquid of dilution
76.25%；Therapeutic effect preventive effect is 63.68%, better than the Propamocarb aqua of comparison medicament 20%.
Claims (9)
1. it is a kind of prevent and treat downy mildew of fruits and vegetables complex micro organism fungicide, it is characterised in that containing two kinds of effective bacterial components, be respectively
Bacillus cercus Bacillus cereus BCJB01, Chinese microorganism strain preservation pipe is preserved on January 30th, 2015
Reason center, deposit number is CGMCC NO.10477；With bacillus megaterium Bacillus megaterium BMJBN02, in
On January 30th, 2015 is preserved in Chinese microorganism strain preservation administrative center, and deposit number is CGMCC NO.10478；
When the complex micro organism fungicide prevents and treats downy mildew of garpe, Bacillus cercus Bacillus cereus BCJB01 and huge
The number of viable ratio of bacillus megateriumBMJBN02 is 1~3:2；It is wax-like during preventing and treating cucumber downy mildew
The viable bacteria of bacillus cereus BCJB01 and bacillus megaterium Bacillus megaterium BMJBN02
Quantity ratio is 1:0.5~2.
2. it is according to claim 1 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide, it is characterised in that the composite microbial
When thing microbial inoculum prevents and treats downy mildew of garpe, Bacillus cercus Bacillus cereus BCJB01 and bacillus megaterium
The number of viable ratio of Bacillus megaterium BMJBN02 is 3:2；During preventing and treating cucumber downy mildew, Bacillus cercus
The number of viable ratio of Bacillus cereus BCJB01 and bacillus megaterium Bacillus megaterium BMJBN02 is
1:2。
3. the preparation method of the complex micro organism fungicide of downy mildew of fruits and vegetables is prevented and treated as claimed in claim 1, it is characterised in that bag
Containing following steps：
(1) culture of seed liquor：By Bacillus cercus Bacillus cereus BCJB01 and bacillus megaterium
Bacillus megaterium BMJBN02 are inoculated into LB liquid medium respectively, and 30 DEG C, 200rpm shaken cultivations 12~18 are small
When, the OD values at 600nm are measured by sampling, returned to zero with nonvaccinated LB liquid medium in continuous mode；
(2) preparation of zymotic fluid：
1. the preparation of Bacillus cercus Bacillus cereus BCJB01 zymotic fluids：When the OD values in step (1) be 0.5~
When 0.8, seed liquor is pressed 1:100 volume ratios add the culture tank equipped with fermentation medium；
2. the preparation of bacillus megaterium Bacillus megaterium BMJBN02 zymotic fluids：When the OD values in step (1) are
When 0.5~0.8, seed liquor is pressed 1:150 volume ratios add the culture tank equipped with fermentation medium；
(3) the mixing compounding of complex microorganism：Take respectively Bacillus cercus Bacillus cereus BCJB01 zymotic fluids and
Bacillus megaterium Bacillus megaterium BMJBN02 fermentating liquid volumes compare 1:1 or 1:3 carry out mixing compounding；
(4) preparation of complex microorganism suspending agent：By step (3) mix the mixing compositional liquor that obtains of compounding and surfactant,
Preservative, suspension aids and thickener are mixed with suspending agent.
4. it is according to claim 3 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
LB liquid medium formula in step (1)：1%NaCl, 1% peptone, 0.5% dusty yeast, pH7.0~7.5.
5. it is according to claim 3 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
The preparation of the 1. Bacillus cercus Bacillus cereus BCJB01 zymotic fluids in step (2), condition of culture is：Culture temperature
30 DEG C of degree, rotating speed 200r/min, fermentation time is 24~36h, and zymotic fluid pH maintains 7.0, dissolved oxygen amount 20%, throughput 5-
7m3/ hour, 0.05~0.1KPa of tank pressure, spore production rate is more than 90%, count plate 8 × 109CFU·mL-1；Fermentation medium
Formula：Corn flour 2%, bean cake powder 1%, dusty yeast 0.3%, CaCO30.3%, MgSO4·7H2O 0.02%, K2HPO4·3H2O
0.2%, KH2PO40.2%, pH7.0~7.2.
6. it is according to claim 3 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
The preparation of the 2. bacillus megaterium Bacillus megaterium BMJBN02 zymotic fluids in step (2), condition of culture is：
30 DEG C of cultivation temperature, rotating speed 220r/min, fermentation time is 36~48h, and zymotic fluid pH maintains 7.0, and dissolved oxygen amount 20% leads to
5~7m of tolerance3/ hour, 0.05~0.1KPa of tank pressure, spore production rate is more than 80%, count plate 5.33 × 109CFU·mL-1；
Fermentative medium formula：Corn flour 2%, bean cake powder 1%, dusty yeast 0.3%, CaCl2 0.1%, MgSO4·7H2O 0.05%,
K2HPO4·3H2O 0.3%, KH2PO40.2%, pH7.0~7.5.
7. it is according to claim 3 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
Suspension aids are ammonium sulfate in step (4), and preservative is glycine, and surfactant is organosilicon BD-3077, and thickener is yellow
Virgin rubber.
8. it is according to claim 7 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
In Bacillus cercus Bacillus cereus BCJB01 zymotic fluids and bacillus megaterium Bacillus in step (4)
In megaterium BMJBN02 zymotic fluid mixing compositional liquors, ammonium sulfate 6%~8%, glycine 2% are added by mass percentage
~5%, BD-3077 0.05%~0.1%, xanthans 0.2%~0.3% is obtained, and count plate is 6.0 in gained suspending agent
~8.0 × 109CFU·mL-1；The consumption of above each component is radix to mix the quality of compositional liquor.
9. it is according to claim 8 preventing and treating downy mildew of fruits and vegetables complex micro organism fungicide preparation method, it is characterised in that
The preparation of step (4) complex microorganism suspending agent is specially：First by the lower addition mixing compositional liquor of ammonium sulfate stirring, ammonium sulfate is treated
All after dissolving, preservative glycine, surfactant organosilicon BD-3077, thickener xanthans are sequentially added under agitation,
Stir.
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| CN105820953B (en) * | 2016-04-06 | 2019-05-31 | 山东省科学院生物研究所 | A kind of screening technique of the efficient biocontrol bacterial strain of downy mildew |
| CN109627103B (en) * | 2019-02-02 | 2021-12-03 | 中盐安徽红四方肥业股份有限公司 | Water-soluble fertilizer for vegetable planting and preparation method and application thereof |
| CN110713954B (en) * | 2019-10-17 | 2022-08-02 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Composite bacillus agent and preparation method and application thereof |
| CN110938559B (en) * | 2019-10-18 | 2022-03-11 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Compound microbial agent for preventing and treating soil-borne diseases, preparation method and application thereof in summer corn cultivation |
| CN110656072B (en) * | 2019-11-19 | 2022-07-19 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Application of bacillus BCJB01 and BMJBN02 in strawberry production |
| CN113652367A (en) * | 2021-07-29 | 2021-11-16 | 海南大学 | Dragon fruit canker bactericide and application thereof |
| CN114158485B (en) * | 2021-12-03 | 2022-12-27 | 安徽省农业科学院农业工程研究所 | Zero-emission straw padding for pig breeding and preparation method thereof |
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