CN108118019A - Bacillus subtilis strain and its application - Google Patents
Bacillus subtilis strain and its application Download PDFInfo
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Abstract
The present invention relates to technical field of microbe application, in particular to a bacillus subtilis strain and its application.It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center to the bacterial strain, and deposit number is：CGMCC NO.15250；The preservation time is：On January 22nd, 2018.The bacterial strain can prevent a variety of true, bacterial diseases, have been supplied at present in soft rot of cabbage, citrus bacterial canker disease, wildfire, tobacco bacterial wilt and bacterial blight of rice.
Description
Technical field
The present invention relates to technical field of microbe application, in particular to a bacillus subtilis strain and its answer
With.
Background technology
Very, bacterial disease is the two class major diseases of growth of harming the crops, and all seriously endangers agricultural production always；Mesh
Before, common prevention is true, the medicament of bacterial disease is mainly chemical pesticide, and general to some disease-controlling effects, right
Ecological environment and agricultural product quality and safety influence very big.Still very lack efficient, wide spectrum, low toxicity, low-residual so far in China
To prevent the biological pesticide based on bacterial disease.
Bacillus subtilis is presently the most the biological prevention and control agent of effective prevention and control plant disease.Foreign countries have multiple withered grass buds
Spore bacillus biocontrol bacterial strain obtains merchandized handling application license.China is successfully developed using bacillus subtilis and puts into life
The commercial preparation of production has sub- precious, hundred anti-(B-908), wheat Fengning (B3), line bent peaceful etc., and control effect is main all more than 50%
It is used for the prevention of proportion of crop planting field diseases.
However, although existing bacillus subtilis formulation is single to plant disease caused by a variety of fungies or bacterium
There is an apparent control effect, but and rare bacterial strain for finding to obtain better effects in disease caused by eubacteria mixes hair.
In addition, the existing method generally use liquid fermentation for preparing bacillus subtilis obtains zymotic fluid, used after pretreatment
Disk centrifugal separator concentrates, and is then spray-dried to obtain original powder；Zymotic fluid is separated by disk centrifugal separator, and heavy-fluid leaves, gently
Liquid discards, and separating rate is fast but yield is low.
In view of this, it is special to propose the present invention.
The content of the invention
The present invention relates to a bacillus subtilis strains, and it is general to be preserved in China Committee for Culture Collection of Microorganisms
Logical microorganism center, deposit number are：CGMCC NO.15250；The preservation time is：On January 22nd, 2018.
The strain Classification system be Bacillus subtilis, Chinese scientific name be bacillus subtilis, the entitled TS- of bacterial strain
55。
The bacillus subtilis strain that above-mentioned process isolates and purifies, 0.7~0.8 × 2~3 microns of individual cells,
Color is uniform.Without pod membrane, peritrichous can move.Gram-positive bacteria, 0.6~0.9 × 1.0~1.5 microns of gemma, ellipse arrives
Column, central or slightly inclined positioned at thalline, thalline does not expand after sporulation.Bacterium colony rough surface is opaque, and dirty white, it passes through
Competitive growth and breeding occupies the mode of living space to prevent the growth of phytopathogen, can quickly form one in plant surface
Layer protective film, makes crops be endangered from pathogen.Multi-functional bacillus subtilis can also secrete antibacterial substance, inhibit germ spore
Son germination and mycelial growth.
During practical application, it is contemplated that it may need the reasons such as transport, it is necessary to be expanded culture and be made
The form of microbial bacterial agent is to expand its application range.
Thus the microbial bacteria of the metabolite containing bacterial strain as described above and/or the bacterial strain is also claimed in the present invention
Agent.
Preferably, microbial bacterial agent as described above, the microbial bacterial agent are solid fungicide, and in the solid fungicide
In, quantity >=10,000,000,000/g of viable bacteria and/or living spores；More preferably >=50,000,000,000/g, >=100,000,000,000/g, >=1,500 hundred million/g or >=
200000000000/g.
Preferably, microbial bacterial agent as described above, the solid fungicide further include solid-state carrier and/or auxiliary agent.
Preferably, microbial bacterial agent as described above, the solid-state carrier include kaolin, precipitated calcium carbonate, diatomite,
One or more in medical stone, calcite, zeolite, white carbon, talcum powder, fine sand and clay.
Preferably, microbial bacterial agent as described above, the auxiliary agent include neopelex, butyl naphthalene sulfonic acids
Sodium, trehalose, glycerine, sodium lignin sulfonate, Negel condensation polymer, niacin, alcohol, buffer salt, sodium chloride, amino acid,
One or more in vitamins, protein, polypeptide, polysaccharide or monose, yeast extract, white carbon, Tea Saponin, defatted milk.
The preparation method of microbial bacterial agent as described above, including：
The strain of the bacillus subtilis is inoculated in fermentation medium and is cultivated and collected zymotic fluid；
Spray drying obtains active compound after the zymotic fluid is mixed with auxiliary agent；
The active compound and solid-state carrier and/or Tea Saponin are mixed to get the solid fungicide.
Preferably, preparation method as described above, in terms of parts by weight, the raw material of the fermentation medium includes：1.8~
3.5 parts of dusty yeast, 0.8~2.0 part of cornstarch, 0.18~0.88 part of calcium carbonate, 0.18~0.88 part of di(2-ethylhexyl)phosphate
Hydrogen potassium, 0.18~0.88 part of ammonium sulfate and 0.018~0.055 part of antifoaming agent；
It is furthermore preferred that in terms of parts by weight, the raw material of the fermentation medium includes：2.0~3.2 parts of dusty yeast, 1.0
~1.6 parts of cornstarch, 0.2~0.6 part of calcium carbonate, 0.2~0.6 part of potassium dihydrogen phosphate, 0.2~0.6 part of sulfuric acid
Ammonium and 0.02~0.04 part of antifoaming agent；
It is furthermore preferred that the dusty yeast is beer yeast powder, and fineness is more than 80 mesh；
It is furthermore preferred that the cornstarch, calcium carbonate, potassium dihydrogen phosphate, ammonium sulfate and antifoaming agent are technical grade.
Preferably, preparation method as described above, the inlet air temperature of the spray drying is 120 DEG C~160 DEG C, goes out wind-warm syndrome
It spends for 70 DEG C~90 DEG C；It is furthermore preferred that the inlet air temperature of the spray drying is 140 DEG C, leaving air temp is 80 DEG C.
According to an aspect of the present invention, the invention further relates to bacterial strains as described above or microbial bacterial agent as described above to exist
Prevent the application in soft rot of cabbage, citrus bacterial canker disease, wildfire, tobacco bacterial wilt or bacterial blight of rice.
Compared with prior art, beneficial effects of the present invention are：
(1) multi-functional bacillus subtilis provided by the present invention is with growth and breeding is fast, fermentation period is short, lower tank spore
The advantages that number height and good control effect.
(2) the preparation method step of multi-functional bacillus subtilis provided by the present invention is simple and convenient to operate, and is by depth
Degree fermentation, ceramic membrane concentration and spray drying obtain multi-functional bacillus subtilis, and preparation efficiency is high, effect is good；Using liquid
Fermentation obtains zymotic fluid, with membrane separation plant concentration after pretreatment, is then spray-dried to obtain original powder；Membrane separation plant is received
Rate is high, can obtain 200,000,000,000 work spore counts/gram original powder.
(3) multi-functional bacillus subtilis provided by the present invention can prevent a variety of true, bacterial diseases, answer at present
With in soft rot of cabbage, citrus bacterial canker disease, wildfire, tobacco bacterial wilt and bacterial blight of rice.
The bacillus subtilis strain (Bacillus subtilis) that the application provides, the entitled TS-55 of bacterial strain are preserved in
China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address are：BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica；The preservation time is：On January 22nd, 2018, deposit number CGMCC NO.15250.
Through collection survival strains are detected as on January 22nd, 2018.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person, the condition suggested according to normal condition or manufacturer carry out.Reagents or instruments used without specified manufacturer is
It can be with conventional products that are commercially available.
Embodiment 1
The present embodiment is related to a kind of preparation method of multi-functional bacillus subtilis microbial agent, comprises the following steps：
1) strain, is prepared, i.e., sterile water is added to make bacteria suspension 10 Kolle flask seeds, is aseptically inoculated into big
In tank；
2), fermentation medium, i.e., by content be 1.8w/v% and fineness >=80 dusty yeasts of mesh, content be 2.0w/v%'s
The PHOSPHORIC ACID TECH.GRADE dihydro that technical grade calcium carbonate that industrial cornstarch, content are 0.18w/v%, content are 0.88w/v%
The technical grade antifoaming agent that the technical grade sulfuric acid ammonium and content that potassium, content are 0.18w/v% are 0.018w/v% is put into body after sterilizing
Product is pH=7.15~7.7 before sterilizing in the tank of 65% volume of fermentation tank；Then throughput is kept within 0~8h this periods
For 1: 0.5, it is that 1: 0.6,11h to tank throughput is put is 1: 0.9 that throughput is kept within 5~12h this periods；0~for 24 hours this
Temperature is 30 DEG C~33 DEG C in the section time, is for 24 hours 29 DEG C~32 DEG C to temperature in tank this period is put, pressure inside the tank is
0.045MPa, speed of agitator 140r/min.
3), post-process, i.e., when fermentation 40 is small or so, microscopy sees in the visual field more than 75% ripe spore, puts tank to tune
Tank to be starched, adjusts pH to 5.5, enters film pressure for 0.3MPa, goes out film pressure 0.2MPa, logical cooling water maintains the temperature at 20 DEG C~30 DEG C,
Cycles of concentration is 5 times.
(inlet air temperature of spray drying is 140 DEG C, and leaving air temp is 80 DEG C, above step for spray drying after adding in auxiliary agent
Spore count can be made as 1.0~10.0 × 1011Original powder)；
Be proportionally added into solid-state carrier and form preparation, add in carrier and Tea Saponin active compound is diluted to 10,000,000,000/gram, auxiliary agent
White carbon is the 10% of gross mass, and Tea Saponin is the 6% of gross mass, remaining is diatomite.
Wherein, auxiliary agent described in step 3) is to be by the mass percent of dry matter in zymotic fluid：2.5% sulfomethylated lignin
Sour sodium, 2.5% dodecyl sodium sulfate, 6% sodium chloride and 0.4% white carbon composition.
The preparation method of the bacillus subtilis, wherein：Spray drying is 140 in inlet air temperature in the step 3)
DEG C, leaving air temp be can be made at 80 DEG C spore count be 200,000,000,000 work spore counts/gram original powder.
Embodiment 2
The present embodiment is related to a kind of preparation method of multi-functional bacillus subtilis microbial agent, comprises the following steps：
1) strain, is prepared, i.e., sterile water is added to make bacteria suspension 10 Kolle flask seeds, is aseptically inoculated into big
In tank；
2), fermentation medium, i.e., by content be 2.2w/v% and fineness >=80 dusty yeasts of mesh, content be 1.5w/v%'s
The PHOSPHORIC ACID TECH.GRADE dihydro that technical grade calcium carbonate that industrial cornstarch, content are 0.5w/v%, content are 0.24w/v%
The technical grade antifoaming agent that the technical grade sulfuric acid ammonium and content that potassium, content are 0.6w/v% are 0.055w/v% is put into volume after sterilizing
In tank for 65% volume of fermentation tank, pH7.15~7.7 before sterilizing；Then it is 1 that throughput is kept within 0~8h this periods:
0.5, it is that 1: 0.6,11h to tank throughput is put is 1: 0.9 that throughput is kept within 5~12h this periods；At 0~this section for 24 hours
Interior temperature is 30 DEG C~33 DEG C, is for 24 hours 29 DEG C~32 DEG C, pressure inside the tank 0.045MPa to temperature in tank this period is put,
Speed of agitator is 140r/min.
3), post-process, i.e., when fermentation 40 is small or so, microscopy sees in the visual field more than 75% ripe spore, puts tank to tune
Tank to be starched, adjusts pH to 5.5, enters film pressure for 0.3MPa, goes out film pressure 0.2MPa, logical cooling water maintains the temperature at 20 DEG C~30 DEG C,
Cycles of concentration is 5 times.
(inlet air temperature of spray drying is 140 DEG C, and leaving air temp is 80 DEG C, above step for spray drying after adding in auxiliary material
Spore count can be made as 1.0~10.0 × 1011Original powder)；
It is proportionally added into auxiliary agent and forms preparation, be proportionally added into auxiliary agent and solid phase carrier forms preparation, add in auxiliary agent by original
Medicine is diluted to 10,000,000,000/gram, auxiliary agent Nekal BX (sodium butylnaphthalenesulfonate) is the 8% of gross mass, and lauryl sodium sulfate is total matter
The 7% of amount, remaining is kaolin support.
Wherein, auxiliary material described in step 3) is to be by the mass percent of dry matter in zymotic fluid：3% lignin sulfonic acid
Sodium or 3% dodecyl sodium sulfate.
The preparation method of the bacillus subtilis, wherein：Spray drying is 140 in inlet air temperature in the step 3)
DEG C, leaving air temp be can be made at 80 DEG C spore count be 200,000,000,000 work spore counts/gram original powder.
The main physical and chemical of multi-functional bacillus subtilis formulation prepared by the present embodiment, it is as shown in the table：
The quality control project and its index of multi-functional bacillus subtilis formulation prepared by the present embodiment, such as following table institute
Show：
The active ingredient of the multi-functional bacillus subtilis (Multi-function Bacillus subtilis) of the present invention
Analysis method is：Using dilution-plate method, bacteria suspension after dilution is quantitatively inoculated in culture medium, treats multi-functional bacillus subtilis
Bacterium grows bacterium colony in culture medium, counts multi-functional bacillus subtilis clump count, with the Unit Weight living spores of determination sample
Number.
The toxicology of the multi-functional bacillus subtilis (Multi-function Bacillus subtilis) of the present invention is：
According to China's pesticide toxicity grading criteria, 100,000,000,000 living spores/gram bacillus subtilis formulation belongs to less toxic fungicide.100000000000 live
Female/male acute oral LD of gemma/gram bacillus subtilis formulation rat505000/ ＞ of ＞, 5000 mg/kgs, acute percutaneous
LD502000/ ＞ of ＞, 2000 mg/kgs, 2000/ ＞ 2000mg/m of Acute inhalation ＞3, low toxicity level.There is slight thorn to rabbit eyes
Swash property, be weak sensitizer to cavy to rabbit no skin irritation.
The prevention of 1 bacterial blight of rice of application examples
Applicant has carried out 2 years four ground field control effectiveness tests using the microbial inoculum being prepared in embodiment 2, as a result takes passages
It is as follows
Determine within 2012 600,750,900 grams/ha of institute's preparation dosage, after third time medicine 14 days preventive effects be respectively 95.64%,
98.33%th, 98.90%, 20% 1500 grams/ha of preventive effects of Yekuzuo wettable powder formulation dosage of comparison medicament are 94.65%.
600,750,900 grams/ha of Guangxi agriculture chemical examination institution preparation dosage, after 3 medicines 14 days preventive effects be respectively 73.30%,
76.88%th, 82.65%, 72% agricultural 312.5 grams/ha of preventive effects of streptomycin sulphate SP preparations dosage of comparison medicament are
74.28%.
600,750,900 grams/ha of agriculture chemical examination institution of Guangdong Province in 2013 preparation dosage is sprayed, and is prevented within 10 days after third time medicine
Effect is respectively 58.26%, 61.89%, 67.65%, 20% 1500 grams of Yekuzuo wettable powder formulation dosage of comparison medicament/public affairs
Hectare preventive effect is 66.64%.600,750,900 grams/ha of Jiangsu Institute for Pesticides's preparation dosage, 10 days preventive effects after second of medicine
Respectively 77.6%, 83.5%, 85.7%, 50% 750 grams/ha of chlorobromide isocyanuric acid SP preparations dosage of comparison medicament, preventive effect
For 84.6%.600,750,900 grams/ha of agriculture chemical examination institution of Fujian Province preparation dosage, 14 days preventive effects are respectively after third time medicine
53.89%th, 61.96%, 66.88%, 20% 1500 grams/ha of preventive effects of Yekuzuo wettable powder formulation dosage of comparison medicament are
60.68%.600,750,900 grams/ha of Guangxi agriculture chemical examination institution preparation dosage, after 2 medicines preventive effect be respectively 64.85%,
70.9%th, 74.88%, 72% agricultural 312.5 grams/ha of preventive effects of streptomycin sulphate SP preparations dosage of comparison medicament are 65.98%.
The prevention of 2 tobacco bacterial wilt of application examples
Applicant has carried out 2 years four ground field control effectiveness tests using the microbial inoculum being prepared in embodiment 2, as a result takes passages
It is as follows：
14 days after 600,750,900 grams/ha of spray 2 medicines of basal part of stem of agriculture chemical examination institution of Guizhou Province in 2013 preparation dosage
Preventive effect is respectively 45.32%, 62.65%, 69.65%, and 72% agricultural streptomycin soluble powder 540mg/kg of comparison medicament prevents
Effect 60.86%.600,750,900 grams/ha of spray stems of Yunnan Prov Academy Of Agricultural Sciences, Agricultural Research Institute Of Environmental Resources preparation dosage
Preventive effect is respectively 52.34%, 61.67%, 65.84% after 3 medicines of base portion, 450 grams of 72% agricultural streptomycin of comparison medicament/public affairs
Hectare, preventive effect 72.250%.14 after 600,750,900 grams/ha of spray 2 medicines of basal part of stem of agriculture chemical examination institution of Henan Province preparation dosage
Its preventive effect is respectively 62.55%, 66.58%, 68.16%, comparison medicament 300,000,000,000/Pseudomonas fluorescence pulvis 8812.5
Gram/ha, preventive effect 65.67%.600,750,900 grams/ha of agriculture chemical examination institution of Shandong Province preparation dosage spray basal part of stem 4 times
7 days preventive effects are respectively 60.12%, 60.97%, 64.85% after medicine, 100,000,000,000 gemma of comparison medicament/gram bacillus subtilis
WP900 grams/ha, preventive effect 63.93%.
10 days after 600,750,900 grams/ha of spray 3 medicines of basal part of stem of agriculture chemical examination institution of Guizhou Province in 2014 preparation dosage
Preventive effect is respectively 56.14%, 66.36%, 68.24%, and 72% agricultural streptomycin soluble powder 720mg/kg of comparison medicament prevents
Effect 74.68%.600,750,900 grams/ha of spray stems of Yunnan Prov Academy Of Agricultural Sciences, Agricultural Research Institute Of Environmental Resources preparation dosage
Preventive effect is respectively 51.68%, 63.66%, 69.06% after 3 medicines of base portion, 450 grams of 72% agricultural streptomycin of comparison medicament/public affairs
Hectare, preventive effect 69.79%.14 after 600,750,900 grams/ha of spray 2 medicines of basal part of stem of agriculture chemical examination institution of Henan Province preparation dosage
Its preventive effect is respectively 58.46%, 64.78%, 65.69%, comparison medicament 300,000,000,000/Pseudomonas fluorescence pulvis 8812.5
Gram/ha, preventive effect 62.90%.600,750,900 grams/ha of agriculture chemical examination institution of Shandong Province preparation dosage spray basal part of stem 3 times
10 days preventive effects are respectively 68.22%, 79.16%, 82.36% after medicine, 1350 grams of 30% nonylphenolsulfonate copper microemulsion of comparison medicament/public affairs
Hectare, preventive effect 78.87%.
The prevention of 3 wildfire of application examples
Applicant has carried out 2 years four ground field control effectiveness tests using the microbial inoculum being prepared in embodiment 2, as a result takes passages
It is as follows：
14 days preventive effects point after agriculture chemical examination institution of Guizhou Province in 2012 preparation dosage 600,750,900 grams/ha of sprayings, 1 medicine
Not Wei 66.93%, 77.68%, 82.69%, 10,000,000,000 living spores of comparison medicament/gram bacillus subtilis wettable powder 750
Gram/ha, preventive effect 76.85%.600,750,900 grams of Yunnan Prov Academy Of Agricultural Sciences, Agricultural Research Institute Of Environmental Resources preparation dosage/
Preventive effect is respectively 54.16%, 64.89%, 71.79% after hectare 3 medicines of spraying, 77% copper calcium sulfate wettable powder of comparison medicament
1250 grams/ha of agent formulation, preventive effect 73.79%.600,750, the 900 grams/ha of sprayings 2 of agriculture chemical examination institution of Henan Province preparation dosage
14 days preventive effects are respectively 72.97%, 75.89%, 77.66% after secondary medicine, the solvable pulvis of 50% chlorobromide isocyanuric acid of comparison medicament
525 grams/ha, preventive effect 73.89%.Agriculture chemical examination institution of Shandong Province preparation dosage 600,750,900 grams/ha of sprayings, 4 medicines
7 days preventive effects are respectively 60.57%, 62.77%, 65.66% afterwards, 100,000,000,000 gemma of comparison medicament/gram bacillus subtilis WP900
Gram/ha, preventive effect 62.36%.
Preventive effect is respectively after agriculture chemical examination institution of Guizhou Province in 2013 preparation dosage 600,750,900 grams/ha of sprayings, 2 medicines
53.65%, 56.87%, 66.45%, 72% agricultural streptomycin soluble powder 720mg/kg preventive effects 74.66% of comparison medicament.
Preventive effect point after Yunnan Prov Academy Of Agricultural Sciences, Agricultural Research Institute Of Environmental Resources preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines
Not Wei 58.78%, 62.58%, 68.68%, 79.09% 1250 grams/ha of copper calcium sulfate wettable powder formulation, preventive effect
71.36%.14 days preventive effects are respectively after agriculture chemical examination institution of Henan Province preparation dosage 600,750,900 grams/ha of sprayings, 2 medicines
65.93%th, 68.89%, 71.89%, 525 grams/ha of the solvable pulvis of 50% chlorobromide isocyanuric acid of comparison medicament, preventive effect is
67.90%.9 days preventive effects are respectively after agriculture chemical examination institution of Shandong Province preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines
78.47%th, 83.79%, 86.56%, 1500 grams/ha of 20% Yekuzuo wettable powder of comparison medicament, preventive effect is
81.55%.
The prevention of 4 citrus bacterial canker disease of application examples
Applicant has carried out 2 years four ground field control effectiveness tests using the microbial inoculum being prepared in embodiment 2, as a result takes passages
It is as follows：
Preventive effect is respectively after 2000 times of agriculture chemical examination institution of Guangdong Province in 2012 liquid, 1500 times of liquid, 1000 times of liquid 3 medicines of spraying
Blade 53.69%, fruit 58.69%, blade 60.17%, fruit 65.89%, blade 68.69%, fruit 71.66%, control
Medicament 77% copper hydroxide wettable powder, 600 times of liquid blades 96.38%, fruit 92.69%.Agriculture chemical examination institution of Fujian Province 2000
Times liquid, 1500 times of liquid, 1000 times of liquid are sprayed 20 days preventive effect difference blades 96.59% after 3 medicines, fruit 92.17%, blade
98.96%, fruit 97.79%, blade 98.89%, fruit 100%, 500 times of 77% copper hydroxide wettable powder of comparison medicament
Liquid, blade 96.48%, fruit 86.79%.2000 times of liquid of Citrus Research Institute Of Chinese Academy Of Agricultural Sciences, 1500 times of liquid, 1000 times of liquid
12 days blade preventive effects are respectively 75.69%, 74.58%, 76.58% after 3 medicines of spraying, and 77% Kocide SD of comparison medicament is wettable
Property pulvis 600 times of liquid preventive effects be 72.69%.Guangxi agriculture chemical examination manages 2000 times of liquid, 1500 times of liquid, 1000 times of liquid sprayings 3
14 days blade preventive effects 65.38% after secondary medicine, fruit preventive effect 62.15%, blade preventive effect 74.06%, fruit preventive effect 71.47%, leaf
Piece preventive effect 76.87%, fruit preventive effect 76.66%, comparison medicament WP500 times of liquid of 77% Kocide SD, blade preventive effect 83.97%,
Fruit preventive effect 81.69%.
20 days preventive effects point after 2000 times of agriculture chemical examination institution of Guangdong Province in 2013 liquid, 1500 times of liquid, 1000 times of liquid 3 medicines of spraying
Not Wei blade 51.86%, fruit 54.58%, blade 58.32%, fruit 62.57%, blade 66.26%, fruit 71.69%,
Comparison medicament 77% copper hydroxide wettable powder, 600 times of liquid blades 78.56%, fruit 81.36%.Agriculture chemical examination institution of Fujian Province
20 days blade preventive effects are respectively 82.38%, 73.78% after 2000 times of liquid, 1500 times of liquid, 1000 times of liquid 3 medicines of spraying,
64.16%, comparison medicament 76% copper hydroxide wettable powder, 500 times of liquid, preventive effect 81.38%.Chinese Academy of Agricultural Sciences's citrus
2000 times of liquid of research institute, 1500 times of liquid, 1000 times of liquid spray fruit preventive effect after 4 medicines be respectively 82.31%, 88.09%,
82.16%, 77% copper hydroxide wettable powder of comparison medicament, 600 times of liquid preventive effects are 87%.Guangxi agriculture chemical examination management institute
20 days blade preventive effects 66.26% after 2000 times of liquid, 1500 times of liquid, 1000 times of liquid 3 medicines of spraying, fruit preventive effect 67.86%, blade
Preventive effect 77.63%, fruit preventive effect 72.96%, blade preventive effect 78.89%, fruit preventive effect 76.44%, 77% hydrogen-oxygen of comparison medicament
Change WP500 times of liquid of copper, blade preventive effect 81.67%, fruit preventive effect 78.35%.
The prevention of 5 soft rot of cabbage of application examples
Applicant has carried out 2 years four ground field control effectiveness tests using the microbial inoculum being prepared in embodiment 2, as a result takes passages
It is as follows：
After agriculture chemical examination institution of Henan Province in 2012 preparation dosage 600,750,900 grams/ha of sprayings, 2 medicines, 15 days preventive effects
Respectively 61.7%, 64.9%, 66.7%, 750 grams/ha of the solvable powder preparation dosage of 50% chlorobromide isocyanuric acid of comparison medicament
Preventive effect is 62.69%.10 days preventive effects point after agriculture chemical examination institution of Shandong Province preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines
Not Wei 75.89%, 82.69%, 86.89%, 20% 1500 grams/ha of Yekuzuo wettable powder formulation amount of comparison medicament, prevent
It imitates as 81.65%.After Vegetable ＆ Flower Inst., Chinese Academy of Agriculture Science's preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines
7 days preventive effects are respectively 61.69%, 73.58%, 85.79%, the comparison medicament 100,000,000,000/agricultural streptomycin sulphate of work spore+72%
150 grams+420 grams/ha of soluble powder agent formulation dosage, preventive effect 72.67%.Heilongjiang Province pesticide management calibrating station preparation is used
10 days preventive effects are respectively 58.88%, 74.60%, 76.44% after amount 600,750,900 grams/ha of sprayings, 2 medicines, comparison medicament
72% agricultural 225 grams/ha of streptomycin sulphate soluble powder agent formulation dosage of bacillus subtilis 100,000,000,000/work spore, preventive effect
For 75.95%.
15 days preventive effects point after agriculture chemical examination institution of Henan Province in 2013 preparation dosage 600,750,900 grams/ha of sprayings, 2 medicines
Not Wei 64.69%, 68.70%, 71.98%, 750 grams of the solvable powder preparation dosage of 50% chlorobromide isocyanuric acid of comparison medicament/public affairs
Hectare preventive effect is 67.8%.10 days preventive effects after agriculture chemical examination institution of Shandong Province preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines
Respectively 63.36%, 70.86%, 78.57%, comparison medicament bacillus subtilis 100,000,000,000/150 gram+72% of work spore is agricultural
225 grams/ha of streptomycin sulphate soluble powder amount of formulation, preventive effect 70.68%.Chinese Academy of Agricultural Sciences's vegetable or flower research
7 days preventive effects are respectively 65.96%, 71.87%, 83.56% after institute's preparation dosage 600,750,900 grams/ha of sprayings, 3 medicines,
72% agricultural 420 grams/ha of streptomycin sulphate soluble powder agent formulation dosage of comparison medicament, preventive effect 74.35%.Heilongjiang Province
After pesticide management calibrating station preparation dosage 600,750,900 grams/ha of sprayings, 2 medicines 10 days preventive effects be respectively 91.88%,
97.93%th, 96.86%, the agricultural streptomycin sulphate soluble powder of comparison medicament bacillus subtilis 100,000,000,000/work spore+72%
225 grams/ha of agent formulation dosage, preventive effect 96.62%.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe is described in detail the present invention with reference to foregoing embodiments, but it will be understood by those of ordinary skill in the art that：Its
It can still modify to the technical solution recorded in foregoing embodiments either to which part or all technical characteristic
Carry out equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is not made to depart from various embodiments of the present invention skill
The scope of art scheme.
Claims (10)
1. bacillus subtilis strain, which is characterized in that it is commonly micro- that it is preserved in China Committee for Culture Collection of Microorganisms
Bio-Centers, deposit number are：CGMCC NO.15250；The preservation time is：22` days in January, 2018.
2. the microbial bacterial agent containing the metabolite for requiring 1 bacterial strain and/or the bacterial strain.
3. microbial bacterial agent according to claim 2, which is characterized in that the microbial bacterial agent is solid fungicide, and
In the solid fungicide, quantity >=10,000,000,000/g of viable bacteria and/or living spores；Preferably >=200,000,000,000/g.
4. microbial bacterial agent according to claim 3, which is characterized in that the solid fungicide further include solid-state carrier and/
Or auxiliary agent.
5. microbial bacterial agent according to claim 4, which is characterized in that the solid-state carrier includes kaolin, lightweight carbon
One or more in sour calcium, diatomite, medical stone, calcite, zeolite, white carbon, talcum powder, fine sand and clay.
6. microbial bacterial agent according to claim 4, which is characterized in that the auxiliary agent include neopelex,
Sodium butylnaphthalenesulfonate, trehalose, glycerine, sodium lignin sulfonate, Negel condensation polymer, niacin, alcohol, buffer salt, chlorination
One kind in sodium, amino acid, vitamins, protein, polypeptide, polysaccharide or monose, yeast extract, white carbon, Tea Saponin, defatted milk
It is or a variety of.
7. the preparation method of claim 3~6 any one of them microbial bacterial agent, which is characterized in that including：
The strain of the bacillus subtilis is inoculated in fermentation medium and is cultivated and collected zymotic fluid；
Spray drying obtains active compound after the zymotic fluid is mixed with auxiliary agent；
The active compound and solid-state carrier and/or auxiliary agent are mixed to get the solid fungicide.
8. preparation method according to claim 7, which is characterized in that in terms of parts by weight, the original of the fermentation medium
Material includes：1.8~3.5 parts of dusty yeast, 0.8~2.0 part of cornstarch, 0.18~0.88 part of calcium carbonate, 0.18~
0.88 part of potassium dihydrogen phosphate, 0.18~0.88 part of ammonium sulfate and 0.018~0.055 part of antifoaming agent.
9. preparation method according to claim 7, which is characterized in that the inlet air temperature of the spray drying for 120 DEG C~
160 DEG C, leaving air temp is 70 DEG C~90 DEG C.
10. bacterial strain described in claim 1 or claim 2~7 any one of them microbial bacterial agent are in the prevention soft corruption of Chinese cabbage
Application in disease, citrus bacterial canker disease, wildfire, tobacco bacterial wilt or bacterial blight of rice.
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| CN108823130A (en) * | 2018-07-05 | 2018-11-16 | 广西大学 | Inhibit plant pathogenetic bacteria GacS/GacA two-component regulator control system biocontrol bacterial strain GacAI and application |
| CN110278965A (en) * | 2019-08-06 | 2019-09-27 | 山东京青农业科技有限公司 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease |
| CN110352987A (en) * | 2019-07-25 | 2019-10-22 | 广东新景象生物工程有限公司 | A kind of prevention and treatment citrus bacterial canker disease new drug and preparation method |
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| CN110604033A (en) * | 2019-11-05 | 2019-12-24 | 隆尧怡东农牧有限公司 | Preparation method and application of matrix for preventing and treating soft rot of vegetables |
| CN112978940A (en) * | 2021-04-22 | 2021-06-18 | 广州市点滴生物科技有限公司 | Organic sewage biological fermentation treating agent and preparation method thereof |
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| CN110352987A (en) * | 2019-07-25 | 2019-10-22 | 广东新景象生物工程有限公司 | A kind of prevention and treatment citrus bacterial canker disease new drug and preparation method |
| CN110278965A (en) * | 2019-08-06 | 2019-09-27 | 山东京青农业科技有限公司 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease |
| CN110373363A (en) * | 2019-08-22 | 2019-10-25 | 福建三炬生物科技股份有限公司 | A kind of concentration microbial inoculum of bacillusmusilaginosiengineering and its preparation method and application |
| CN110604033A (en) * | 2019-11-05 | 2019-12-24 | 隆尧怡东农牧有限公司 | Preparation method and application of matrix for preventing and treating soft rot of vegetables |
| CN110604033B (en) * | 2019-11-05 | 2021-08-24 | 隆尧怡东农牧有限公司 | Preparation method and application of matrix for preventing and treating soft rot of vegetables |
| CN112978940A (en) * | 2021-04-22 | 2021-06-18 | 广州市点滴生物科技有限公司 | Organic sewage biological fermentation treating agent and preparation method thereof |
| CN112978940B (en) * | 2021-04-22 | 2021-07-16 | 广州市点滴生物科技有限公司 | Organic sewage biological fermentation treating agent and preparation method thereof |
| CN113234646A (en) * | 2021-07-12 | 2021-08-10 | 北京新正和农业科技有限公司 | Bacillus subtilis microbial inoculum and preparation method and application thereof |
| CN113234646B (en) * | 2021-07-12 | 2021-09-24 | 北京新正和农业科技有限公司 | Bacillus subtilis microbial inoculum and preparation method and application thereof |
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