CN109805032A - A kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent and preparation method thereof inhibiting bacterial disease - Google Patents
A kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent and preparation method thereof inhibiting bacterial disease Download PDFInfo
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Abstract
The present invention provides a kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent and preparation method thereof for inhibiting bacterial disease, is grouped as by the group of following mass parts: 2~6 parts of bacillus preparation, 1 part of S-Ethyl ethylthio sulfonate.It is that bacillus is carried out to rejuvenation of spawn and activation processing in LB culture medium when preparation；Triangular flask culture medium, seed tank culture base and fermentation tank culture medium are carried out disinfection processing respectively；Strain inoculated is cultivated into 20h into triangular flask culture medium；The strain inoculated of acquisition is cultivated into 16h into seed tank culture base again；Then the strain inoculated of acquisition is cultivated to 28h into fermentation tank culture medium to get bacillus preparation is arrived；Bacillus preparation is mixed in proportion with S-Ethyl ethylthio sulfonate to get the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease is arrived.Microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of the invention have it is easy to use, it is time saving and energy saving, can be used in mixed way with liquid fertilizer, antibiotic, on microbial inoculum activity without influence, have the advantages that comprehensive benefit is good.
Description
Technical field
The invention belongs to the Prevention Technique fields of citrus disease, and in particular to a kind of bacillus is combined with chemical bacteriostatic agent
The microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent and preparation method thereof of shared inhibition bacterial disease.
Background technique
Antimicrobial spectrum, bacteriostasis and mechanism of different biocontrol bacterial strains etc. have larger difference.Biocontrol Bacillus is repeatedly trying
The ability for being found to have and inhibiting a variety of pathogenic bacterias being widely present in nature is tested, it is more friendly to environment and the mankind, because
This, by the favor of domestic and international plant protection and Study on Environmental Protection person.
China has carried out a large amount of research to the bacillus with Biocontrol Effect at present.And it is anti-in bacillus subtilis
It controls and has had reached advanced international standard in the application study of plant disease.But the currently known biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease
That there are still storage lives is short, not salt tolerant, intolerant to osmotic pressure, chemical bacteriostatic agent have deactivation to it, cannot be total with chemical bacteriostatic agent
With the disadvantages of control efficiency in, resistance difference and use is poor, slow effect.
Currently, country advocates ecologic planting energetically, develop sustainable green agriculture, therefore one in the prevention and treatment of citrus disease
Slightly biological biological and ecological methods to prevent plant disease, pests, and erosion agent is come into being, and at most, but these preparations have slow effect, steady to the stronger bacillus application of resistance more
Qualitative difference, it is weak in plant definite value ability, due Antagonistic Fungi quantity cannot be formed, to inhibit the ability of pathogenic bacteria weak, is prevented and treated
Effect is unstable.Most of bacillus are sensitive to chemical sterilization, bacteriostatic agent, cannot use simultaneously, some to be applied to anti-cure the disease
Harmful chemical residue influences the growth and breeding of biocontrol microorganisms also larger.Therefore microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent is during controlling disease
By various limitations.
Summary of the invention
In order to overcome the deficiencies of the prior art, the present invention provides a kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent and its system for inhibiting bacterial disease
Preparation Method, energy prevention and treatment citrus illness, while the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent is easy to use, can mix with liquid fertilizer and antibiotic makes
With.
The present invention is realized by following technical proposal: a kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent inhibiting bacterial disease, by following matter
The group of amount part is grouped as:
2~6 parts of bacillus preparation, 1 part of S-Ethyl ethylthio sulfonate；
Wherein, bacillus preparation is made of every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings.
The bacillus living is the bacillus for being named as Bacillus DY-3.Routinely pass through ultraviolet light and sulphur
Diethyl phthalate complex mutation, separation, screening, domestication and obtain, be chemically-resistant, antibiotic-resistant, salt tolerant, bacterial strain resistant to high osmotic pressure,
This kind of bacterial strain is suitable for the prevention and treatment of the illnesss such as bacterial canker, black spot, black rot, angular leaf spot, and the present invention is exactly by this kind
Bacterial strain carried out in conjunction with chemical bacteriostatic agent through special formulation using.
The fermentation tank culture medium is made of the component of following mass parts:
94~96 parts of sterile ultrapure waters, 0.5~1 part of glucose, 1~3 part of molasses, 0.1~0.4 part of starch, 0.3~0.8
Part bean powder, 0.3~0.8 part of fish meal, 0.3~0.8 part of peptone, 0.02~0.06 part of potassium dihydrogen phosphate, 0.01~0.03 part of nothing
Water manganese sulfate, 0.01~0.03 part of seven water manganese sulfate.
Another object of the present invention is to provide a kind of preparation method of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease, packets
Include following steps:
Step 1: bacillus is subjected to rejuvenation of spawn and activation processing in LB culture medium；
Step 2: by triangular flask culture medium, seed tank culture base and fermentation tank culture medium: being not less than 0.1MPa, temperature in pressure
Under conditions of degree is not less than 121 DEG C, carry out disinfection processing respectively；
Step 3: the bacterial strain that step 1 processing obtains being seeded in the triangular flask culture medium of step 2 with oese, in temperature
32 DEG C, revolving speed be 200r/min shaking table in cultivate 20h；Again by the seed tank culture base of the strain inoculated of acquisition to step 2,
16h is cultivated under the conditions of revolving speed 200r/min, ventilatory capacity 0.4v/Vmin, filling pressure 0.02Mpa, 32 DEG C of temperature；Then by acquisition
Strain inoculated in revolving speed 160r/min, ventilatory capacity 0.4v/Vmin, fills pressure 0.02Mpa, temperature into the fermentation tank culture medium of step 2
28h is cultivated under the conditions of 32 DEG C of degree, is made in every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings to get the bacillus system of arriving
Agent；
Step 4: by bacillus preparation that step 3 obtains and S-Ethyl ethylthio sulfonate (2~6) in mass ratio: 1 mix to get
To the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease.
The rejuvenation of spawn and activation processing of the step 1 comprise the steps of:
Step S1: take the Bacillus strain in preservation state spare；
Step S2: by LB culture medium after the sterilizing of conventional high temperature autoclave, LB culture medium is fallen on superclean bench
Enter 9 centimetres of sterile petri dish, every culture dish about 15mL；
Step S3: after the solidification of LB culture medium, plate is drawn with the bacillus kind lawn of oese picking preservation；
Step S4: being horizontally arranged more than half an hour, and lawn is waited to be adsorbed on LB culture medium；
Step S5: plate is inverted in 37 DEG C of insulating boxs and is cultivated 30 hours；
Step S6: observing the Bacillus of culture, selects healthy and strong bacterium colony, selects part of colony inoculation to newly
LB culture medium in cultivate, repeat 2-3 time, to obtain well-grown bacterial strain, that is, complete rejuvenation of spawn and activation processing.
Triangular flask culture medium in the step 2 is made of the component of following mass parts:
97~98 parts of sterile ultrapure waters, 0.8~1.2 part of glucose, 0.8~1.2 part of peptone, 0.02~0.04 part of phosphoric acid
Potassium dihydrogen, 0.01~0.03 part of anhydrous manganous sulfate, 0.3~0.8 portion of beef extract, 0.01~0.03 part of epsom salt, 0.01~
0.02 part of sodium chloride adjusts pH to 6.8~7.2 with sodium hydroxide.
Seed tank culture base and fermentation tank culture medium in the step 2 are made of the component of following mass parts:
94~96 parts of sterile ultrapure waters, 0.5~1 part of glucose, 1~3 part of molasses, 0.1~0.4 part of starch, 0.3~0.8
Part bean powder, 0.3~0.8 part of fish meal, 0.3~0.8 part of peptone, 0.02~0.06 part of potassium dihydrogen phosphate, 0.01~0.03 part of nothing
Water manganese sulfate, 0.01~0.03 part of seven water manganese sulfate.
Gained inhibits the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease when in use, is sprayed after diluting it by 1:200 times
Mist is sprayed in whole plant, is sprayed through 2-3 times, disease can be made obviously even to be eliminated by control.
Gained inhibits the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease when in use, is that 1:300 is administered to soil by volume by it
In earth, through the flora microbial action in biological prevention and control agent, constantly discharges various nutrients and utilized for plant absorption, and microbial bacteria
Group constructs new microecosystem balance in the micro-ecological environment around root system of plant, thoroughly overcomes and uses chemicals merely
Treatment is not thorough, disadvantage easy to recur.
The beneficial effects of the present invention are the method combined using two kinds of components can accomplish prevention and treatment simultaneously, right
Citrus plant diseases accomplish effectively to control, and S-Ethyl ethylthio sulfonate is mainly used in the treatment of a large amount of diseases, and bacillus preparation is mainly used
Prevention and treatment before disease occurs or after disease treatment.Crop surface can be directly sprayed on after dilution, the defence to the disease of plant
Play good effect.Solve the problems, such as the prevention and treatment of citrus illness, at the same the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent have it is easy to use,
It is time saving and energy saving, it can be used in mixed way with liquid fertilizer, antibiotic, on microbial inoculum activity without influence, have the advantages that comprehensive benefit is good.
Gained of the invention inhibits the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease under the closed normal temperature state that do not break a seal, and 12 months
Survival rate still keeps 90% or more afterwards, and the biological prevention and control agent of the prior art can only save 6 months.In the inorganic of concentration 30w/w%
Survival rate is up to 95% in salting liquid, and the biological prevention and control agent of the prior art is only resistant to the inorganic salt solution of 20w/w%.General chemistry suppression
Microbial inoculum or antibiotic on bacillus preparation growth and breeding of the invention without influence (such as kasugarnycin, zinc thiazole), and it is conventional
Ordinary preparation cannot be shared with chemobiotic.The present invention, which compares conventional bacterial strain, has stronger resistance.
Specific embodiment
Below with reference to embodiment, the invention will be further described.
Embodiment 1
Step 1: bacillus (being named as Bacillus DY-3) is carried out at rejuvenation of spawn and activation in LB culture medium
Reason:
Step S1: take the Bacillus strain in preservation state spare；
Step S2: by LB culture medium after the sterilizing of conventional high temperature autoclave, LB culture medium is fallen on superclean bench
Enter 9 centimetres of sterile petri dish, every culture dish about 15mL；
Step S3: after the solidification of LB culture medium, plate is drawn with the bacillus kind lawn of oese picking preservation；
Step S4: being horizontally arranged more than half an hour, and lawn is waited to be adsorbed on LB culture medium；
Step S5: plate is inverted in 37 DEG C of insulating boxs and is cultivated 30 hours；
Step S6: observing the Bacillus of culture, selects healthy and strong bacterium colony, selects part of colony inoculation to newly
LB culture medium in cultivate, repeat 2-3 time, to obtain well-grown bacterial strain, that is, complete rejuvenation of spawn and activation processing.
Step 2: by triangular flask culture medium, seed tank culture base and fermentation tank culture medium: being not less than 0.1MPa, temperature in pressure
Under conditions of degree is not less than 121 DEG C, carry out disinfection processing respectively；
Wherein, triangular flask culture medium is made of the component of following mass parts:
97 parts of sterile ultrapure waters, 1 part of glucose, 1 part of peptone, 0.03 part of potassium dihydrogen phosphate, 0.02 part of anhydrous manganous sulfate,
0.5 portion of beef extract, 0.02 part of epsom salt, 0.01 part of sodium chloride adjust pH to 6.8~7.2 with sodium hydroxide；
Seed tank culture base and fermentation tank culture medium are made of the component of following mass parts:
96 parts of sterile ultrapure waters, 0.8 part of glucose, 2 parts of molasses, 0.2 part of starch, 0.5 part of bean powder, 0.5 part of fish meal, 0.5
Part peptone, 0.04 part of potassium dihydrogen phosphate, 0.02 part of anhydrous manganous sulfate, 0.02 part of seven water manganese sulfate.
Step 3: the bacterial strain that step 1 processing obtains being seeded in the triangular flask culture medium of step 2 with oese, in temperature
32 DEG C, revolving speed be 200r/min shaking table in cultivate 20h；Again by the seed tank culture base of the strain inoculated of acquisition to step 2,
16h is cultivated under the conditions of revolving speed 200r/min, ventilatory capacity 0.4v/Vmin, filling pressure 0.02Mpa, 32 DEG C of temperature；Then by acquisition
Strain inoculated in revolving speed 160r/min, ventilatory capacity 0.4v/Vmin, fills pressure 0.02Mpa, temperature into the fermentation tank culture medium of step 2
28h is cultivated under the conditions of 32 DEG C of degree, is made in every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings to get the bacillus system of arriving
Agent；
Step 4: bacillus preparation that step 3 obtains being mixed with S-Ethyl ethylthio sulfonate 5:1 in mass ratio to get to inhibiting
The microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease.
Embodiment 2
Step 1: bacillus (being named as Bacillus DY-3) is carried out at rejuvenation of spawn and activation in LB culture medium
Reason:
Step S1: take the Bacillus strain in preservation state spare；
Step S2: by LB culture medium after the sterilizing of conventional high temperature autoclave, LB culture medium is fallen on superclean bench
Enter 9 centimetres of sterile petri dish, every culture dish about 15mL；
Step S3: after the solidification of LB culture medium, plate is drawn with the bacillus kind lawn of oese picking preservation；
Step S4: being horizontally arranged more than half an hour, and lawn is waited to be adsorbed on LB culture medium；
Step S5: plate is inverted in 37 DEG C of insulating boxs and is cultivated 30 hours；
Step S6: observing the Bacillus of culture, selects healthy and strong bacterium colony, selects part of colony inoculation to newly
LB culture medium in cultivate, repeat 2-3 time, to obtain well-grown bacterial strain, that is, complete rejuvenation of spawn and activation processing.
Step 2: by triangular flask culture medium, seed tank culture base and fermentation tank culture medium: in pressure 0.3MPa, temperature 130
Under conditions of DEG C, carry out disinfection processing respectively；
Wherein, triangular flask culture medium is made of the component of following mass parts:
98 parts of sterile ultrapure waters, 1.2 parts of glucose, 1.2 parts of peptones, 0.04 part of potassium dihydrogen phosphate, 0.03 part of anhydrous sulphur
Sour manganese, 0.8 portion of beef extract, 0.03 part of epsom salt, 0.02 part of sodium chloride adjust pH to 6.8~7.2 with sodium hydroxide；
Seed tank culture base and fermentation tank culture medium are made of the component of following mass parts:
94 parts of sterile ultrapure waters, 1 part of glucose, 3 parts of molasses, 0.4 part of starch, 0.8 part of bean powder, 0.8 part of fish meal, 0.8 part
Peptone, 0.06 part of potassium dihydrogen phosphate, 0.03 part of anhydrous manganous sulfate, 0.03 part of seven water manganese sulfate.
Step 3: the bacterial strain that step 1 processing obtains being seeded in the triangular flask culture medium of step 2 with oese, in temperature
32 DEG C, revolving speed be 200r/min shaking table in cultivate 20h；Again by the seed tank culture base of the strain inoculated of acquisition to step 2,
16h is cultivated under the conditions of revolving speed 200r/min, ventilatory capacity 0.4v/Vmin, filling pressure 0.02Mpa, 32 DEG C of temperature；Then by acquisition
Strain inoculated in revolving speed 160r/min, ventilatory capacity 0.4v/Vmin, fills pressure 0.02Mpa, temperature into the fermentation tank culture medium of step 2
28h is cultivated under the conditions of 32 DEG C of degree, is made in every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings to get the bacillus system of arriving
Agent；
Step 4: bacillus preparation that step 3 obtains being mixed with S-Ethyl ethylthio sulfonate 2:1 in mass ratio to get to inhibiting
The microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease.
Embodiment 3
Step 1: bacillus (being named as Bacillus DY-3) is carried out at rejuvenation of spawn and activation in LB culture medium
Reason:
Step S1: take the Bacillus strain in preservation state spare；
Step S2: by LB culture medium after the sterilizing of conventional high temperature autoclave, LB culture medium is fallen on superclean bench
Enter 9 centimetres of sterile petri dish, every culture dish about 15mL；
Step S3: after the solidification of LB culture medium, plate is drawn with the bacillus kind lawn of oese picking preservation；
Step S4: being horizontally arranged more than half an hour, and lawn is waited to be adsorbed on LB culture medium；
Step S5: plate is inverted in 37 DEG C of insulating boxs and is cultivated 30 hours；
Step S6: observing the Bacillus of culture, selects healthy and strong bacterium colony, selects part of colony inoculation to newly
LB culture medium in cultivate, repeat 2-3 time, to obtain well-grown bacterial strain, that is, complete rejuvenation of spawn and activation processing.
Step 2: by triangular flask culture medium, seed tank culture base and fermentation tank culture medium: in pressure 0.5MPa, temperature 135
Under conditions of DEG C, carry out disinfection processing respectively；
Wherein, triangular flask culture medium is made of the component of following mass parts:
98 parts of sterile ultrapure waters, 0.8 part of glucose, 0.8 part of peptone, 0.02 part of potassium dihydrogen phosphate, 0.01 part of anhydrous sulphur
Sour manganese, 0.3 portion of beef extract, 0.01 part of epsom salt, 0.01 part of sodium chloride adjust pH to 6.8~7.2 with sodium hydroxide；
Seed tank culture base and fermentation tank culture medium are made of the component of following mass parts:
95 parts of sterile ultrapure waters, 0.5 part of glucose, 1 part of molasses, 0.1 part of starch, 0.3 part of bean powder, 0.3 part of fish meal, 0.3
Part peptone, 0.02 part of potassium dihydrogen phosphate, 0.01 part of anhydrous manganous sulfate, 0.01 part of seven water manganese sulfate.
Step 3: the bacterial strain that step 1 processing obtains being seeded in the triangular flask culture medium of step 2 with oese, in temperature
32 DEG C, revolving speed be 200r/min shaking table in cultivate 20h；Again by the seed tank culture base of the strain inoculated of acquisition to step 2,
16h is cultivated under the conditions of revolving speed 200r/min, ventilatory capacity 0.4v/Vmin, filling pressure 0.02Mpa, 32 DEG C of temperature；Then by acquisition
Strain inoculated in revolving speed 160r/min, ventilatory capacity 0.4v/Vmin, fills pressure 0.02Mpa, temperature into the fermentation tank culture medium of step 2
28h is cultivated under the conditions of 32 DEG C of degree, is made in every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings to get the bacillus system of arriving
Agent；
Step 4: bacillus preparation that step 3 obtains being mixed with S-Ethyl ethylthio sulfonate 6:1 in mass ratio to get to inhibiting
The microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease.
Comparative example 1: with the embodiment of the present invention 1, bacillus preparation is used only, is not incorporated S-Ethyl ethylthio sulfonate.
Comparative example 2: with the embodiment of the present invention 1, S-Ethyl ethylthio sulfonate is used only, is not incorporated bacillus preparation.
Comparative example 3: with the embodiment of the present invention 1, the proportion of bacillus preparation and S-Ethyl ethylthio sulfonate is 1:4.
Comparative example 4: with the embodiment of the present invention 1, bacillus DY-3 is only replaced with into solution starch gemma bar
Bacterium.
Comparative example 5: with the embodiment of the present invention 1, bacillus DY-3 is only replaced with into bacillus subtilis.
Comparative example 6: with the embodiment of the present invention 1, only omitting step 1-3, directly using bacillus DY-3 and
S-Ethyl ethylthio sulfonate mixing.
Above-described embodiment and comparative example are used in citrus plantation, sprayed by spraying after each example is diluted by 1:200 times
In in whole plant, acquired results are as follows:
| Product storage life | Product survival rate | Resistance | Control rate | |
| Embodiment 1 | 12 months | 95.12% | By force | 96.36% |
| Embodiment 2 | 12 months | 95.33% | By force | 96.87% |
| Embodiment 3 | 12 months | 95.07% | By force | 95.41% |
| Comparative example 1 | 12 months | 96.78% | By force | 73.45% |
| Comparative example 2 | 12 months | -- | -- | 76.59% |
| Comparative example 3 | 12 months | 91.56% | By force | 89.81% |
| Comparative example 4 | 6 months | 12.49% | Difference | 66.32% |
| Comparative example 5 | 6 months | 32.56% | Difference | 69.75% |
| Comparative example 6 | 9 months | 85.69% | Generally | 87.2% |
In the disease control of citrus, the bacillus with biological and ecological methods to prevent plant disease, pests, and erosion function is applied in plant plant protection protection,
But not by large-scale popularization application, main cause is exactly that effect is unstable, slow effect, and citrus bacterial canker disease outbreak period bacillus is not
Can effective definite value, be not achieved prevention and treatment require.The present invention overcomes disadvantage mentioned above, in citrus bacterial canker disease outbreak period bacillus system
Agent is treated with S-Ethyl ethylthio sulfonate mixture, and S-Ethyl ethylthio sulfonate kills rapidly pathogen, creates growing environment, gemma bar for bacillus definite value
Bacterium succeeds, and definite value is sustainable to avoid infecting again for pathogen.It is only prevented and treated, is kept away with bacillus preparation in citrus disease initial phase
Exempt from disease outburst, reduces drug cost.If conventional bacillus shares with S-Ethyl ethylthio sulfonate, S-Ethyl ethylthio sulfonate can cause bacillus to lose
Living, bacillus is unable to definite value, does not have the effect that long-term pre- disease prevention occurs.
Using the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent provided by the invention for inhibiting bacterial disease, is sprayed through 2-3 times, disease can be made bright
It is aobvious even to be eliminated by control.
Claims (9)
1. a kind of microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease, it is characterised in that be grouped as by the group of following mass parts:
2~6 parts of bacillus preparation, 1 part of S-Ethyl ethylthio sulfonate；
Wherein, bacillus preparation is made of every milliliter of fermentation tank culture medium containing 1,000,000,000 bacillus livings.
2. the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent according to claim 1 for inhibiting bacterial disease, it is characterised in that: the bacillus
Viable bacteria is the bacillus for being named as Bacillus DY-3.
3. the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent according to claim 1 for inhibiting bacterial disease, it is characterised in that: the fermentor training
Base is supported to be made of the component of following mass parts:
94~96 parts of sterile ultrapure waters, 0.5~1 part of glucose, 1~3 part of molasses, 0.1~0.4 part of starch, 0.3~0.8 part of beans
Powder, 0.3~0.8 part of fish meal, 0.3~0.8 part of peptone, 0.02~0.06 part of potassium dihydrogen phosphate, 0.01~0.03 part of anhydrous sulphur
Sour manganese, 0.01~0.03 part of seven water manganese sulfate.
4. a kind of preparation method for the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent for inhibiting bacterial disease, it is characterised in that the following steps are included:
Step 1: bacillus is subjected to rejuvenation of spawn and activation processing in LB culture medium；
Step 2: by triangular flask culture medium, seed tank culture base and fermentation tank culture medium: being not less than 0.1MPa, temperature not in pressure
Under conditions of 121 DEG C, carry out disinfection processing respectively；
Step 3: the strain inoculated that step 1 processing is obtained is in 32 DEG C of temperature, revolving speed into the triangular flask culture medium of step 2
20h is cultivated in the shaking table of 200r/min；Again by the seed tank culture base of the strain inoculated of acquisition to step 2, in revolving speed 200r/
Min, ventilatory capacity 0.4v/Vmin, it fills pressure 0.02Mpa, cultivate 16h under the conditions of 32 DEG C of temperature；Then the strain inoculated of acquisition is arrived
In the fermentation tank culture medium of step 2, in revolving speed 160r/min, ventilatory capacity 0.4v/Vmin, pressure 32 DEG C of 0.02Mpa, temperature conditions are filled
Lower culture 28h makes to contain 1,000,000,000 bacillus livings in every milliliter of fermentation tank culture medium to get bacillus preparation is arrived；
Step 4: by bacillus preparation that step 3 obtains and S-Ethyl ethylthio sulfonate (2~6) in mass ratio: 1 mixes to get to suppression
The microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of bacterial disease processed.
5. the preparation method according to claim 4, it is characterised in that: the rejuvenation of spawn and activation processing packet of the step 1
Containing following steps:
Step S1: take the Bacillus strain in preservation state spare；
Step S2: by LB culture medium after conventional sterilant, LB culture medium is poured into sterile petri dish；
Step S3: after the solidification of LB culture medium, the bacillus kind lawn of picking preservation draws plate；
Step S4: being horizontally arranged more than half an hour, and lawn is waited to be adsorbed on LB culture medium；
Step S5: plate is inverted in 37 DEG C and is cultivated 30 hours；
Step S6: observing the Bacillus of culture, selects healthy and strong bacterium colony, selects part of colony inoculation to new LB
It cultivates, repeats 2-3 times in culture medium, to obtain well-grown bacterial strain, that is, complete rejuvenation of spawn and activation processing.
6. the preparation method according to claim 4, it is characterised in that: the triangular flask culture medium in the step 2 is by following
The component of mass parts is constituted:
97~98 parts of sterile ultrapure waters, 0.8~1.2 part of glucose, 0.8~1.2 part of peptone, 0.02~0.04 part of biphosphate
Potassium, 0.01~0.03 part of anhydrous manganous sulfate, 0.3~0.8 portion of beef extract, 0.01~0.03 part of epsom salt, 0.01~0.02
Part sodium chloride adjusts pH to 6.8~7.2 with sodium hydroxide.
7. the preparation method according to claim 4, it is characterised in that: seed tank culture base and fermentation in the step 2
Tank culture medium is made of the component of following mass parts:
94~96 parts of sterile ultrapure waters, 0.5~1 part of glucose, 1~3 part of molasses, 0.1~0.4 part of starch, 0.3~0.8 part of beans
Powder, 0.3~0.8 part of fish meal, 0.3~0.8 part of peptone, 0.02~0.06 part of potassium dihydrogen phosphate, 0.01~0.03 part of anhydrous sulphur
Sour manganese, 0.01~0.03 part of seven water manganese sulfate.
8. the preparation method according to claim 4, it is characterised in that: the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of gained inhibition bacterial disease
It when in use, is sprayed in whole plant by spraying after diluting it by 1:200 times.
9. the preparation method according to claim 4, it is characterised in that: the microorganism biological and ecological methods to prevent plant disease, pests, and erosion agent of gained inhibition bacterial disease
It when in use, is that 1:300 is administered in soil by volume by it.
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