CN110278965A - A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease - Google Patents
A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease Download PDFInfo
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- CN110278965A CN110278965A CN201910721017.9A CN201910721017A CN110278965A CN 110278965 A CN110278965 A CN 110278965A CN 201910721017 A CN201910721017 A CN 201910721017A CN 110278965 A CN110278965 A CN 110278965A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/005—Following a specific plan, e.g. pattern
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention discloses a kind of microbial bacterial agents for preventing and treating citrus bacterial canker disease, are related to microorganisms technical field, the constituent and its mass fraction of microbial-bacterial fertilizer are as follows: microbial bacteria 5-40%, adsorbent 30-80%, wetting agent 1-5%, suspending agent 2-10%；The microbial bacteria of microbial bacterial agent is bacillus subtilis, living bacteria count >=30,000,000,000 cfu/g of microbial bacteria.The present invention also provides a kind of preparation method of microbial bacterial agent for preventing and treating citrus bacterial canker disease, the preparation of microbial bacterial agent includes the preparation of bacillus subtilis microbial agent and the preparation of microbial bacterial agent.The microbial bacterial agent of prevention and treatment citrus bacterial canker disease provided by the invention is used in mixed way with copper agent, can effectively extend the lasting period, reduces copper agent usage amount, the more efficient generation for preventing citrus bacterial canker disease, infected plant, safety coefficient are higher for healing.
Description
Technical field
The present invention relates to microorganisms technical field, in particular to a kind of microbial bacterial agent and its system for preventing and treating citrus bacterial canker disease
Preparation Method.
Background technique
With the rapid development of citrus plantation industry, cultivated area rapid expansion causes citrus bacterial canker disease cause of disease in orchard
Bacterium radix increases year by year, and then citrus bacterial canker disease is caused to be on the rise, and will cause treetop ulcer lesion, and blade is largely sent out
Disease, fruit rind deformity, ulcer seriously affect fruit appearance and mouthfeel.Citrus bacterial canker disease is the important disease of citrus
One of, pathogenic bacteria Strain differentiation is complicated, disease incidence is high, propagation is fast, host range is wide, seriously affects the yield and product of citrus
Matter becomes the significant threat of Citrus industry development.
Existing market prevents citrus bacterial canker disease usually using copper agent, but largely brings a system using copper agent for a long time
Column problem, such as: copper deposition reduces citrus photosynthesis, aggravates red spider outburst, pathogenic bacteria resistance to drugs increase and citrus
Heavy metals exceeding standard etc..Bio-control method is compared with other methods as a kind of emerging control measure, have safely, effectively,
Lasting feature especially can be avoided chemical prevention bring a series of problems, good, environmental-friendly to disease-controlling effect,
It is able to maintain the ecological balance, effective component is that natural products is degradable, therefore the research of bio-control method becomes a kind of and necessarily becomes
Gesture.
Research shows that Bacillus cercus can efficiently prevent and treat citrus bacterial canker disease, control efficiency is up to 2017 Nian Wangchao et al.
65.46% (107586735 A of CN), however, since there are safety issues for Bacillus cercus, for eating industrial crops
On, people is edible to cause to poison by food, and poisoner's symptom is abdominal pain, vomiting, diarrhea etc..On March 20th, 2010, the Ministry of Agriculture, China
It stopped micro- life of the strain according to " microbial manure bio-safety general technology criterion " (NY 1109-2006) relevant regulations
The registration of object fertilizer.
Summary of the invention
According to technological deficiency existing in the prior art, the technical problems to be solved by the present invention are: providing a kind of prevention and treatment
Microbial bacterial agent of citrus bacterial canker disease and preparation method thereof reduces pesticide residue and endangers caused by plant growth, solves simultaneously
Using side effect brought by Bacillus cercus microbial inoculum, the disease incidence of citrus bacterial canker disease is reduced, improves the yield and product of citrus
Matter.
In order to realize technical purpose of the invention, a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease of the present invention and its preparation
The technical solution of method is:
A kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease, the constituent and its mass fraction of the microbial bacterial agent
Are as follows: microbial bacteria 5-40%, adsorbent 30-80%, wetting agent 1-5%, suspending agent 2-10%.
As a further improvement of the present invention, the microbial bacteria is bacillus subtilis, the bacillus subtilis
(Bacillus subtilis), abbreviation JQ-001 bacterial strain are preserved in Chinese microorganism strain preservation on November 09th, 2015
Administration committee's common micro-organisms center, depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, deposit number are
CGMCC No.11632。
As a further improvement of the present invention, the living bacteria count of the microbial bacteria >=30,000,000,000 cfu/g.
As a further improvement of the present invention, the adsorbent be kaolin, diatomite, bentonite, light calcium one kind or
It is a variety of.
As a further improvement of the present invention, the wetting agent be pull open powder, K12, NNO, EFW it is one or more.
As a further improvement of the present invention, the suspending agent is sodium lignin sulfonate, D425, sodium carboxymethylcellulose
It is one or more.
As a further improvement of the present invention, the dosage form of the microbial bacterial agent is wettable powder.
As a further improvement of the present invention, the preparation method of the microbial bacterial agent includes the following steps:
(1) actication of culture: the bacillus subtilis of cryo-conservation is inoculated on beef-protein medium,
24-36h is cultivated under 36-38 DEG C of constant temperature；
(2) preparation of primary seed solution: preparing beef extract-peptone Shake flask medium, is dispensed using 500mL triangular flask, dress
Liquid measure 200mL, the bacillus subtilis after high pressure sterilization after access activation, 36-38 DEG C of temperature, revolving speed 200-260rpm/
22-26h is cultivated under the conditions of min；
(3) preparation of secondary seed solution: preparing seed tank culture base, and the primary seed solution that step (2) is obtained is pressed
The inoculum concentration of 3-5% is respectively connected to seeding tank, the seeding tank liquid amount 60%, and 36-38 DEG C of temperature, revolving speed 180-220rpm/
20-26h is cultivated under the conditions of min, ventilation ratio 1:1；The formula of the seed tank culture base is as follows: peptone 0.8-1.2%, yeast
Extract 0.4-0.6%, sodium chloride 0.8-1.2%；
(4) large-scale industry fermented and cultured: preparation fermentation tank culture medium, the secondary seed solution that step (3) is obtained,
It is respectively connected to large-scale industry fermentor by the 5-10% inoculum concentration of large-scale industry fermentor used medium volume ratio, it is described
Fermentation tank capacity 60%, 36-38 DEG C of temperature, revolving speed 180-260rpm/min, culture is to generating bud under the conditions of ventilatory capacity ratio 1:1
Born of the same parents complete fermentation, obtain bacillus subtilis fermentation liquor；The fermentation tank culture based formulas is as follows: bean cake powder 3.5-4.5%,
Glucose 4.5-5.5%, potassium dihydrogen phosphate 0.2-0.25%；
(5) it the preparation of bacillus subtilis microbial agent: in the bacillus subtilis fermentation liquor obtained to step (4), presses
Fermentation liquid dry matter weight ratio is that 1:1 addition adsorbent is adsorbed, and is then directly spray-dried；
(6) detection of the bacillus subtilis microbial agent bacteria containing amount: using microbial inoculum after dilution-plate method detection spray drying
Bacteria containing amount；
(7) preparation of the microbial bacterial agent: the gemma bacillus agent that step (5) is obtained is by weight addition
60% adsorbent, 2% wetting agent, 8% suspending agent are uniformly mixed, and airslide disintegrating mill, fineness control is added in the material mixed
325-400 mesh detects the technical indicators such as wettable powder bacteria containing amount, suspension, wetability, obtains after up to standard described micro- after crushing
Bacteria agent.
As a further improvement of the present invention, the microbial bacterial agent and copper agent compounding application, for preventing and treating Citrus
Plant canker, specific Dressing date and dosage are as follows:
(1) spring tip lamina 1 week or so, located again with 1000 times of the microbial bacteria dilution agent+copper agent 1000
Reason, pouring root 1 time, foliage-spray 2 times (interval 15-20 days)；
(2) long to 2-3 centimetres of summer shoot when, carried out with 1000 times+described 1000 times of copper agent of the microbial bacteria dilution agent
Processing, pouring root 1 time, foliage-spray 2 times (interval 15-20 days)；
(3) long to 2-3 centimetres of autumn growth when, carried out with 1000 times+described 1000 times of copper agent of the microbial bacteria dilution agent
Processing, pouring root 1 time, foliage-spray 2 times (interval 15-20 days).
Compared with prior art, the beneficial effects of the present invention are:
The microbial bacterial agent of prevention and treatment citrus bacterial canker disease provided by the invention has with short production cycle, active high, at low cost
Advantage, is conducive to industrialized production, and the bacillus subtilis of application is a kind of salt tolerant, heat-resisting, aerobic and have existence extensively suitable
Should be able to power bacterium, antagonism multiple pathogenic microorganisms, the further diffusion of pre- disease prevention, diseases prevention, improve citrus resistance, promote
Raw, conditioning soil etc. plays the role of vital, and bacillus subtilis is hidden without there is safety in use
Suffer from problem.The microbial bacterial agent is used in mixed way with copper agent, can effectively extend the lasting period, reduces copper agent usage amount, more
It is effectively prevent the generation of citrus bacterial canker disease, infected plant, safety coefficient are higher for healing.
Specific embodiment
Bacillus subtilis (Bacillus subtilis) provided by the invention, abbreviation JQ-001 bacterial strain is in 2015
November 09 was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution address: Beijing
The institute of Chaoyang District North Star West Road 1, deposit number are CGMCC No.11632.
To be further elaborated to beneficial effect of the present invention, following specific embodiments have been carried out, special instruction, below
Specific embodiment is intended to better understand the present invention, is not restricted to the scope of the present invention.Experimental method in following embodiments,
It unless otherwise specified, is conventional method.Experimental material used in following embodiments is unless otherwise specified shop purchase
Routine biochemistry preparation.Quantitative test in following embodiment is respectively provided with and repeats to test three times, and results are averaged.
The preparation of embodiment 1, microbial bacterial agent
1, a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease, the constituent and its mass fraction of microbial bacterial agent are as follows:
Microbial bacteria 30%, adsorbent 60%, wetting agent 2%, suspending agent 8%.
2, microbial bacteria is bacillus subtilis, bacillus subtilis (Bacillus subtilis), abbreviation JQ-001 bacterium
Strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution on November 09th, 2015
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, deposit number are CGMCC No.11632.
The identification related content of bacillus subtilis JQ-001 bacterial strain is as follows:
The identification (CGMCC No.11632) of bacillus subtilis JQ-001 bacterial strain
(1) Microbiological Characteristics: bacterial strain is under NA culture medium, 37 DEG C of environment, and after cultivating 12-18h, bacterium colony rough surface is not
Transparent, dirty white or yellowish, flat dry matt, edge is irregular, not chromogenesis.Bacterial strain is rod-shaped, has peritrichous,
Gram's staining is positive, aerobic.
(2) molecular biological characteristic: the 16SrDNA extension increasing sequence of JQ-001 bacterial strain is compared in ncbi database, with
Bacillus subtilis homology is 99.38%, and morphological feature, physiological and biochemical property in conjunction with JQ-001 bacterial strain determine
JQ-001 bacterial strain belongs to bacillus subtilis Bacillus subtilis.
3, the living bacteria count of microbial bacteria >=30,000,000,000 cfu/g.
4, adsorbent be kaolin, diatomite, bentonite, light calcium it is one or more.
5, wetting agent be pull open powder, K12, NNO, EFW it is one or more.
6, suspending agent be sodium lignin sulfonate, D425, sodium carboxymethylcellulose it is one or more.
7, the dosage form of microbial bacterial agent is wettable powder.
8, the preparation method of microbial bacterial agent includes the following steps:
(1) actication of culture: the bacillus subtilis of cryo-conservation is inoculated on beef-protein medium, 36-38
24-36h is cultivated under DEG C constant temperature；
(2) preparation of primary seed solution: preparing beef extract-peptone Shake flask medium, is dispensed using 500mL triangular flask, dress
Liquid measure 200mL, the bacillus subtilis after high pressure sterilization after access activation, 36-38 DEG C of temperature, revolving speed 200-260rpm/min item
22-26h is cultivated under part；
(3) seed tank culture base, the primary seed solution that step (2) is obtained, by 3- the preparation of secondary seed solution: are prepared
5% inoculum concentration is respectively connected to seeding tank, and seeding tank liquid amount 60%, leads to by 36-38 DEG C of temperature, revolving speed 180-220rpm/min
20-26h is cultivated under the conditions of gas ratio 1:1；The formula of seed tank culture base is as follows: peptone 0.8-1.2%, yeast extract 0.4-
0.6%, sodium chloride 0.8-1.2%；
(4) fermentation tank culture medium, the secondary seed solution that step (3) is obtained, by big large-scale industry fermented and cultured: are prepared
The 5-10% inoculum concentration of technical scale fermentor used medium volume ratio is respectively connected to large-scale industry fermentor, ferments canned
Liquid measure 60%, 36-38 DEG C of temperature, revolving speed 180-260rpm/min, culture is completed to brood cell is generated under the conditions of ventilatory capacity ratio 1:1
Fermentation, obtains bacillus subtilis fermentation liquor；Fermentation tank culture based formulas is as follows: bean cake powder 3.5-4.5%, glucose 4.5-
5.5%, potassium dihydrogen phosphate 0.2-0.25%；
(5) preparation of bacillus subtilis microbial agent: in the bacillus subtilis fermentation liquor obtained to step (4), by fermentation
Liquid dry matter weight ratio is that 1:1 addition adsorbent is adsorbed, and is then directly spray-dried；
(6) detection of bacillus subtilis microbial agent bacteria containing amount: microbial inoculum contains after being spray-dried using dilution-plate method detection
Bacterium amount；
(7) preparation of microbial bacterial agent: the gemma bacillus agent that step (5) is obtained is adsorbed by weight being added 60%
Agent, 2% wetting agent, 8% suspending agent are uniformly mixed, and airslide disintegrating mill are added in the material mixed, fineness controls 325-400
Mesh detects the technical indicators such as wettable powder bacteria containing amount, suspension, wetability, obtains microbial bacterial agent after up to standard after crushing.
9, microbial bacterial agent and copper agent compounding application, for preventing and treating citrus plant canker, specific Dressing date and
Dosage is as follows:
(1) it spring tip lamina 1 week or so, is handled again with 1000 times+copper agent of microbial bacteria dilution agent 1000, pouring root 1
It is secondary, foliage-spray 2 times (interval 15-20 days)；
(2) long to 2-3 centimetres of summer shoot when, handled again with 1000 times+copper agent of microbial bacteria dilution agent 1000, pouring root
1 time, foliage-spray 2 times (interval 15-20 days)；
(3) long to 2-3 centimetres of autumn growth when, handled again with 1000 times+copper agent of microbial bacteria dilution agent 1000, pouring root
1 time, foliage-spray 2 times (interval 15-20 days).
The influence of embodiment 2, microbial bacterial agent to fertile mandarin orange citrus bacterial canker disease
1, reagent agent
The microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention.
2, for studying object and controlling object
For studying object: fertile mandarin orange, the age of tree 5 years；
Controlling object: citrus bacterial canker disease.
3, test site
Test site: the county Meng Man, autonomous prefecture, Xishuangbanna of Yunnan province the Dai nationality.
4, field experiment
1. experimental design
3 processing of test setting, each processing choose 6 plants of fruit tree, and processing I is blank control；The 500 times of liquid copper of processing II
Preparation is handled；Processing III is handled with 1000 times of liquid+1000 times of liquid copper agents of microbial bacterial agent of the present invention.
Citrus concentrates 3 applications, and timing node is respectively as follows: in mid or late Febuary, 2018, in mid-June, 2018, and 2018 9
The middle ten days and the last ten days moon.
It carries out pouring root 1 time, foliage-spray 2 times, time interval 15-20 days every time.
2. test result
Influence result such as table 1 of the microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention to fertile mandarin orange citrus bacterial canker disease
It is shown.
Influence of 1 microbial bacterial agent of table to fertile mandarin orange citrus bacterial canker disease
| Processing | Disease refers to preventive effect % | Kg/ plants of average product |
| Handle I (blank control) | 0 | 29.7 |
| Processing II | 47.0 | 30.2 |
| Processing III | 84.7 | 46.7 |
As it can be seen from table 1 the microbial bacterial agent that the present invention prevents and treats citrus bacterial canker disease has preferably fertile mandarin orange citrus bacterial canker disease
Disease refer to preventive effect.
The influence of embodiment 3, microbial bacterial agent to granulated sugar tangerine citrus bacterial canker disease
1, reagent agent
The microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention.
2, for studying object and controlling object
For studying object: granulated sugar tangerine, the age of tree 5 years；
Controlling object: citrus bacterial canker disease.
3, test site
Test site: Guangxi Zhuang Autonomous Region Bose City Xilin County.
4, field experiment
1. experimental design
3 processing of test setting, each processing choose 6 plants of fruit tree, and processing I is blank control；The 500 times of liquid copper of processing II
Preparation is handled；Processing III is handled with 1000 times of liquid+1000 times of liquid copper agents of microbial bacterial agent of the present invention.
Citrus concentrates 3 applications, and timing node is respectively as follows: in mid or late Febuary, 2018, in mid-June, 2018, and 2018 9
The middle ten days and the last ten days moon.
It carries out pouring root 1 time, foliage-spray 2 times, time interval 15-20 days every time.
2. test result
The microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention to the influence result of granulated sugar tangerine citrus bacterial canker disease such as
Shown in table 2.
Influence of 2 microbial bacterial agent of table to granulated sugar tangerine citrus bacterial canker disease
| Processing | Disease refers to preventive effect % | Kg/ plants of average product |
| Handle I (blank control) | 0 | 57.8 |
| Processing II | 42.1 | 59.0 |
| Processing III | 79.9 | 86.3 |
From table 2 it can be seen that the present invention prevent and treat citrus bacterial canker disease microbial bacterial agent to granulated sugar tangerine citrus bacterial canker disease have compared with
Good disease refers to preventive effect.
The influence of embodiment 4, microbial bacterial agent to crystal sugar orange citrus bacterial canker disease
1, reagent agent
The microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention.
2, for studying object and controlling object
For studying object: crystal sugar orange, the age of tree 3 years；
Controlling object: citrus bacterial canker disease.
3, test site
Test site: Hunan Province Chenzhou City Yongxing County.
4, field experiment
1. experimental design
3 processing of test setting, each processing choose 6 plants of fruit tree, and processing I is blank control；The 500 times of liquid copper of processing II
Preparation is handled；Processing III is handled with 1000 times of liquid+1000 times of liquid copper agents of microbial bacterial agent of the present invention.
Citrus concentrates 3 applications, and timing node is respectively as follows: in mid or late Febuary, 2018, in mid-June, 2018, and 2018 9
The middle ten days and the last ten days moon.
It carries out pouring root 1 time, foliage-spray 2 times, time interval 15-20 days every time.
2. test result
Influence result of the microbial bacterial agent of prevention and treatment citrus bacterial canker disease prepared by the present invention to crystal sugar orange citrus bacterial canker disease is such as
Shown in table 3.
Influence of 3 microbial bacterial agent of table to crystal sugar orange citrus bacterial canker disease
| Processing | Disease refers to preventive effect % | Kg/ plants of average product |
| Handle I (blank control) | 0 | 27.3 |
| Processing II | 44.7 | 29.4 |
| Processing III | 82.0 | 41.8 |
From table 3 it can be seen that the present invention prevent and treat citrus bacterial canker disease microbial bacterial agent to crystal sugar orange citrus bacterial canker disease have compared with
Good disease refers to preventive effect.
Microbial bacterial agent prepared by the present invention and copper agent compounding application can be effectively reduced citrus bacterial canker disease pathogen
It infects and propagates, to reduce blade and fruit scab number, reduce leaf abscission, and the disease-resistant lasting period is longer；Meanwhile the present invention
Microbial bacterial agent can dramatically increase the percentage of fertile fruit of citrus, to improve the yield and quality of citrus.
The above is only preferable embodiment of the invention, is not intended to limit the present invention in any form, Ren Hewei
It is detached from technical solution of the present invention content, according to the technical essence of the invention to any simple modification, transformation made by above example
Material equivalent variations and modification, all of which are still within the scope of the technical scheme of the invention.
Claims (9)
1. it is a kind of prevent and treat citrus bacterial canker disease microbial bacterial agent, it is characterised in that: the constituent of the microbial bacterial agent and its
Mass fraction are as follows: microbial bacteria 5-40%, adsorbent 30-80%, wetting agent 1-5%, suspending agent 2-10%.
2. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 1, it is characterised in that: the microorganism
Bacterium is bacillus subtilis, and the bacillus subtilis (Bacillus subtilis), abbreviation JQ-001 bacterial strain is in 2015
November 09 was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution address: Beijing
The institute of Chaoyang District North Star West Road 1, deposit number are CGMCC No.11632.
3. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 2, it is characterised in that: the microorganism
The living bacteria count of bacterium >=30,000,000,000 cfu/g.
4. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 3, it is characterised in that: the adsorbent
For kaolin, diatomite, bentonite, light calcium it is one or more.
5. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 4, it is characterised in that: the wetting agent
For pull open powder, K12, NNO, EFW it is one or more.
6. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 5, it is characterised in that: the suspending agent
For sodium lignin sulfonate, D425, sodium carboxymethylcellulose it is one or more.
7. a kind of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 6, it is characterised in that: the microorganism
The dosage form of microbial inoculum is wettable powder.
8. a kind of preparation method of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 1, it is characterised in that:
The preparation method of the microbial bacterial agent includes the following steps:
(1) actication of culture: the bacillus subtilis of cryo-conservation is inoculated on beef-protein medium, 36-38
24-36h is cultivated under DEG C constant temperature；
(2) preparation of primary seed solution: preparing beef extract-peptone Shake flask medium, is dispensed using 500mL triangular flask, liquid amount
200mL, the bacillus subtilis after high pressure sterilization after access activation, 36-38 DEG C of temperature, revolving speed 200-260rpm/min item
22-26h is cultivated under part；
(3) seed tank culture base, the primary seed solution that step (2) is obtained, by 3- the preparation of secondary seed solution: are prepared
5% inoculum concentration is respectively connected to seeding tank, the seeding tank liquid amount 60%, and 36-38 DEG C of temperature, revolving speed 180-220rpm/
20-26h is cultivated under the conditions of min, ventilation ratio 1:1；The formula of the seed tank culture base is as follows: peptone 0.8-1.2%, yeast
Extract 0.4-0.6%, sodium chloride 0.8-1.2%；
(4) fermentation tank culture medium, the secondary seed solution that step (3) is obtained, by big large-scale industry fermented and cultured: are prepared
The 5-10% inoculum concentration of technical scale fermentor used medium volume ratio is respectively connected to large-scale industry fermentor, the fermentation
Canned liquid measure 60%, 36-38 DEG C of temperature, revolving speed 180-260rpm/min, under the conditions of ventilatory capacity ratio 1:1 culture to generating brood cell,
Fermentation is completed, bacillus subtilis fermentation liquor is obtained；The fermentation tank culture based formulas is as follows: bean cake powder 3.5-4.5%, grape
Sugared 4.5-5.5%, potassium dihydrogen phosphate 0.2-0.25%；
(5) preparation of bacillus subtilis microbial agent: in the bacillus subtilis fermentation liquor obtained to step (4), by fermentation
Liquid dry matter weight ratio is that 1:1 addition adsorbent is adsorbed, and is then directly spray-dried；
(6) detection of the bacillus subtilis microbial agent bacteria containing amount: microbial inoculum contains after being spray-dried using dilution-plate method detection
Bacterium amount；
(7) preparation of the microbial bacterial agent: the gemma bacillus agent that step (5) is obtained is inhaled by weight being added 60%
Attached dose, 2% wetting agent, 8% suspending agent is uniformly mixed, and airslide disintegrating mill is added in the material mixed, fineness controls 325-400
Mesh detects the technical indicators such as wettable powder bacteria containing amount, suspension, wetability, the microbial bacteria is obtained after up to standard after crushing
Agent.
9. a kind of application of microbial bacterial agent for preventing and treating citrus bacterial canker disease according to claim 1, it is characterised in that: described
Microbial bacterial agent and copper agent compounding application, for preventing and treating citrus plant canker, specific Dressing date and dosage are as follows:
(1) it spring tip lamina 1 week or so, is handled, is filled again with 1000 times of the microbial bacteria dilution agent+copper agent 1000
Root 1 time, foliage-spray 2 times (interval 15-20 days)；
(2) long to 2-3 centimetres of summer shoot when, handled again with 1000 times of the microbial bacteria dilution agent+copper agent 1000,
Pouring root 1 time, foliage-spray 2 times (interval 15-20 days)；
(3) long to 2-3 centimetres of autumn growth when, handled again with 1000 times of the microbial bacteria dilution agent+copper agent 1000,
Pouring root 1 time, foliage-spray 2 times (interval 15-20 days).
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910721017.9A CN110278965A (en) | 2019-08-06 | 2019-08-06 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease |
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| CN201910721017.9A CN110278965A (en) | 2019-08-06 | 2019-08-06 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating citrus bacterial canker disease |
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| CN111657301A (en) * | 2020-05-27 | 2020-09-15 | 西安工程大学 | Wettable plant rhizosphere microorganism growth promoting powder and preparation method thereof |
| CN112042448A (en) * | 2020-09-17 | 2020-12-08 | 保定市科绿丰生化科技有限公司 | Citrus canker control method based on bacillus subtilis DnKas |
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