CN103013839B - Pasty biocontrol preparation for cucumber fusarium wilt, and preparation method, application and special strain thereof - Google Patents
Pasty biocontrol preparation for cucumber fusarium wilt, and preparation method, application and special strain thereof Download PDFInfo
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Abstract
The invention relates to a pasty biocontrol preparation for cucumber fusarium wilt, and a preparation method, application and a special strain thereof. The classification designation of the special strain is Trichoderma asperellum GY20, and the collection number is CGMCC NO:4899. The pasty biocontrol preparation provided by the invention takes hyphae and chlamydospores of the Trichoderma asperellum GY20 as active components of the preparation. According to the invention, the pasty biocontrol preparation has the biggest advantage of stable product quality, and can be stored for 18 months at room temperature and 3 years under refrigeration conditions; the pasty biocontrol preparation has favorable control effect on cucumber fusarium wilt; and waste water, waste gas and waste residues are never generated, thereby being beneficial to environmental protection.
Description
Technical field
The present invention relates to microorganism field, in particular to a kind of cucumber fusarium axysporum paste biological prevention and control agent,
Preparation method, application and special strain therefore thereof.
Background technology
Cucumber is a kind of global important vegetable.In recent years, because cucumber cultivation area expands year by year, large-scale south vegetable north transportation in addition, cucumber is become can the important vegetables of year-round supply, in " vegetable basket project ", account for critical role.But; along with the mass-producing of cucumber production, Base environment, specialization; and relevant variation of ecology and environment; disease problem on cucumber production seems very outstanding; occurrence degree is on the rise; especially the harm of the soil-borne disease such as cucumber fusarium axysporum is larger, has had a strong impact on the yield and quality of gourd vegetable crop.
Cucumber fusarium axysporum
(Cucumber Fusarium wilt)also known as dead arm, wilt disease, be by Fusarium oxysporum (
fusarium axysporumf.sp.
cucumerinum) cause typical soil-borne disease.In the area of cultivated cucumber kind all the year round, cucumber fusarium axysporum sickness rate is about 10% ~ 20%, and serious can reach more than 50%.Estimate according to experienced vegetable grower, the underproduction rate of generally falling ill in early stage of gathering is 70% ~ 100% of sickness rate, and mid-term is 30% ~ 50%, and the underproduction rate growing latter stage is then less, also may cause total crop failure time serious.Therefore, cucumber fusarium axysporum has become the serious development of obstruction green cucumber industry and the important disease increased income of peasant.
Production generally adopt the method such as breeding resistant variety, graft seedling growth, chemical control reduce blight to producing the loss caused.Existing cucumber variety, even if minority kind has certain anti-, resistance to sick ability (as middle peasant No. 5, Tianjin spring No. 5 etc.), fundamentally can not solve the generation of disease; Graft technology difficulty is comparatively large, is difficult to spread; Chemical control as simply, controlling way fast and effectively, obtains the favor of people, but there is no effective agricultural chemicals and can prevent and treat this kind of disease, and the existing chemicals of life-time service easily develops immunity to drugs, and easily can cause severe contamination to environment.Therefore, by seeking novel diseases prevention, means of curing the disease, reaching the harm effectively controlling cucumber fusarium axysporum, having become problem in the urgent need to address in production practice.
Microbial pesticide comes from nature, has the features such as environment compatibility is good, the lasting period is long, safety low-poison, becomes one of important environmental protection approach of various fruit diseases control.But up to now, utilize the genetically engineered bacteria strain of fungi and paste biological prevention and control agent high-efficiency prevention and control cucumber fusarium axysporum thereof then to rarely have report.
Summary of the invention
The object of the invention is to solve above-mentioned the deficiencies in the prior art and problem, a kind of genetically engineered bacteria strain for preventing and treating high-efficiency prevention and control cucumber fusarium axysporum, paste biological prevention and control agent and its preparation method and application containing this bacterial strain are provided.
In order to realize object of the present invention, the invention provides a strain gene engineering bacteria strain, its Classification And Nomenclature be trichoderma asperellum (
trichoderma asperellum) GY20, its deposit number is: CGMCC NO:4899.
Present invention also offers a kind of paste biological prevention and control agent of cucumber fusarium axysporum, wherein, the activeconstituents of described preparation be above-mentioned mention described trichoderma asperellum (
trichoderma asperellum) mycelia of GY20 and chlamydospore.
Further, described mycelia and chlamydosporic ratio, determined by following cultural method:
By grown on potato dextrose agar trichoderma asperellum (
trichoderma asperellum) GY20 spore is with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, by 5%(V/V) inoculum size is inoculated in potato dextrose broth, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato grape liquid nutrient medium shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain.
Preferably, the weight part content of each component of fermentation culture in described fermentor tank is: beet liquid 380-420, magnesium sulfate heptahydrate 3-5, potassium primary phosphate 38-42; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 300-400 rev/min, leavening temperature 25-27 degree Celsius, fermentation time 48 hours, pH 7.0-7.2; The condition of described shaking culture is 140-160 rev/min, 25-27 degree Celsius; Condition centrifugal after described fermentation culture is 7500-8500 rev/min, centrifugal 10 minutes.
Top condition is: the fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2.
Fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2.
Preferably, the condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Condition centrifugal after described fermentation culture be 8000 revs/min centrifugal 10 minutes.
Preferably, described paste biological prevention and control agent also comprises the copper ion inhibitor of 0.001 ~ 0.05%.
Preferably, described paste biological prevention and control agent also comprises the Zulkovsky starch of 5 ~ 10%.
Preferably, described Zulkovsky starch is potato starch.
Present invention also offers a kind of method preparing the paste biological prevention and control agent of above-mentioned cucumber fusarium axysporum, wherein, described method comprises:
1) by grown on potato dextrose agar trichoderma asperellum (
trichoderma asperellum) GY20 spore is with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, by 5%(V/V) inoculum size is inoculated in potato dextrose broth, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato grape liquid nutrient medium shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain.
2) adjust ph: the pH value of above-mentioned mycelia and chlamydospore mixture is adjusted to 3.0 ~ 4.0;
3) obtained paste biological prevention and control agent: to described step 2) add in the mixture that obtains 0.001 ~ 0.05% copper ion inhibitor, finally add the Zulkovsky starch of 5 ~ 10%, stir.
Preferably, the weight part content of each component of fermentation culture in described fermentor tank is: beet liquid 380-420, magnesium sulfate heptahydrate 3-5, potassium primary phosphate 38-42; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 300-400 rev/min, leavening temperature 25-27 degree Celsius, fermentation time 48 hours, pH 7.0-7.2; The condition of described shaking culture is 140-160 rev/min, 25-27 degree Celsius; Condition centrifugal after described fermentation culture is 7500-8500 rev/min, centrifugal 10 minutes.
Top condition is: the condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2; Condition centrifugal after described fermentation culture be 8000 revs/min centrifugal 10 minutes; Described Zulkovsky starch is potato starch.
Moreover, present invention also offers described genetically engineered bacteria strain trichoderma asperellum (
trichoderma asperellum) application in control cucumber fusarium axysporum of GY20 and obtained paste biological prevention and control agent.
The present invention for prevent and treat cucumber fusarium axysporum containing genetically engineered bacteria strain trichoderma asperellum (
trichoderma asperellum) the paste biological prevention and control agent of GY20 compared with existing " viride " paste biological prevention and control agent, have the following advantages:
1, paste biological prevention and control agent of the present invention is made up of active ingredient and auxiliary material, active ingredient be trichoderma asperellum (
trichoderma asperellum) GY20 mycelia and chlamydospore, its chlamydospore is dispersed in Trichoderma silk, the paste biological prevention and control agent constant product quality that it is prepared into, can room temperature preservation 18 months, preserves 3 years under refrigerated condition;
2, paste biological prevention and control agent of the present invention has good preventive effect to cucumber fusarium axysporum: greenhouse test result shows, GY20 all can reach more than 90% to the prevention effect of cucumber fusarium axysporum, is significantly higher than " viride " viride " paste biological prevention and control agent is to the prevention effect 58.40% of cucumber fusarium axysporum;
3, nuisanceless: trichoderma asperellum of the present invention (
trichoderma asperellum) GY20 can the fungi of safe release, therefore a series of negative environment effects that can not bring because of the use of agricultural chemicals, the use simultaneously containing the mould paste biological prevention and control agent of this wood greatly reduces the amount of application of chemical pesticide in cucumber production, any waste water and gas waste residue can not be produced, for the control of cucumber harmless boilogical provides Reliable guarantee, benefit modern agriculture with high efficiency;
4, cost is lower: trichoderma asperellum of the present invention (
trichoderma asperellum) GY20 and paste biological prevention and control agent production cost very low, enterprise obtain profit margin very big.。
Embodiment
Biomaterial trichoderma asperellum of the present invention (
trichoderma asperellum) preservation date of GY20 is on May 25th, 2011, depositary institution's full name is China Microbiological biological inoculum preservation management committee's common micro-organisms center, being called for short: CGMCC, address is: Beijing. Institute of Microorganism, Academia Sinica, its deposit number is: CGMCC NO:4899.
Described trichoderma asperellum (
trichoderma asperellum) GY20 has following feature: cultivate 4d under 25 DEG C of dark conditions, colony diameter 53-55mm, white mycelium, cotton-shaped, luxuriant; Conidium structure produces in a large number, and Chan Bao district is uniformly distributed, greyish-green; Bottle stalk formula produces spore, and conidium is oval, subsphaeroidal, and there is spinelet on surface.Trichoderma has effect to multiple important plant pathogenic fungi, as pythium spp, Verticillium, sickle-like bacteria, Helminthosporium sp. bacterium, Alternariaspp, rhizoctonia, botrytis etc.
A paste biological prevention and control agent for cucumber fusarium axysporum, wherein, the activeconstituents of described preparation for trichoderma asperellum described in claim 1 (
trichoderma asperellum) mycelia of GY20 and chlamydospore.
Wherein, described mycelia and chlamydosporic ratio, determined by following cultural method:
By grown on potato dextrose agar trichoderma asperellum (
trichoderma asperellum) GY20 spore is with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, by 5%(V/V) inoculum size is inoculated in potato dextrose broth, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato grape liquid nutrient medium shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain.
Wherein, the fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2; The condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Condition centrifugal after described fermentation culture be 8000 revs/min centrifugal 10 minutes.
Further, the pH value of described paste biological prevention and control agent is 3.0 ~ 4.0, and preferred pH value is 3.5.
Further, described paste biological prevention and control agent also comprises the copper ion inhibitor of 0.001 ~ 0.05%, and preferred described paste biological prevention and control agent comprises the copper ion inhibitor of 0.001 ~ 0.05%, the preferably MDA-5 of 0.01%.
Further, described paste biological prevention and control agent also comprises the Zulkovsky starch of 5 ~ 10%.Preferably, described paste biological prevention and control agent also comprises the Zulkovsky starch of 5 ~ 10%.Preferably, described Zulkovsky starch is the potato starch of 8%.
On the other hand, the present invention protects a kind of method preparing the paste biological prevention and control agent of cucumber fusarium axysporum described above, and wherein, described method comprises:
1) by grown on potato dextrose agar trichoderma asperellum (
trichoderma asperellum) GY20 spore is with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, by 5%(V/V) inoculum size is inoculated in potato dextrose broth, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato grape liquid nutrient medium shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain.
2) adjust ph: the pH value of above-mentioned mycelia and chlamydospore mixture is adjusted to 3.0 ~ 4.0;
3) obtained paste biological prevention and control agent: to described step 2) add in the mixture that obtains 0.001 ~ 0.05% copper ion inhibitor, finally add the Zulkovsky starch of 5 ~ 10%, stir.
Wherein, the condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2; Condition centrifugal after described fermentation culture be 8000 revs/min centrifugal 10 minutes; Described Zulkovsky starch is potato starch.
Particularly, paste biological prevention and control agent prepared by control cucumber fusarium axysporum bacterial strain GY20 of the present invention, its preparation method comprises the following steps:
(A) acquisition of bacterium liquid
With aseptic inoculation cutter picking in advance at the upper cultured GY20 spore of potato dextrose agar (PDA), saving into spore content with aseptic washing downward is 10
5the inoculation liquid of spore/mL, by 5%(V/V) to be inoculated in the volume that 200mL potato grape liquid nutrient medium (PD) is housed be in the triangular flask of 750 mL to inoculum size, in 26 DEG C, shaking culture 48 h under 150 rpm/min conditions, it is in the triangular flask of 2000 mL that upper step gained nutrient solution 200 mL equal portions are inoculated in 4 volumes that 500mL PD nutrient solution is housed, shaking culture 48 h under 150 rpm, 26 DEG C of conditions; The 2000mL nutrient solution of gained is inoculated in the fermentor tank (east, Zhenjiang GUS-30) that the volume that 20L nutrient solution is housed is 30L, 20L fermentation culture liquid hold-up is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g, arranging fermentation condition is: dissolved oxygen 100%, stirring velocity is 350 rpm, leavening temperature 26 DEG C, fermentation time 48 h, pH 7.0-7.2.
(B) paste biological prevention and control agent complete processing
The fermented liquid fermented is processed into paste biological prevention and control agent as follows: first by fermented liquid centrifugal 10min under 8000rpm/min condition, obtain mycelia and chlamydospore mixture, with salt acid for adjusting pH to 3.0 ~ 4.0, and add the copper ion inhibitor of 0.001 ~ 0.05%, finally add the Zulkovsky starch of 5 ~ 10%, stir and namely made paste biological prevention and control agent.
Preferably, after regulating mycelia and chlamydospore mixture pH to 3.0 ~ 4.0 with hydrochloric acid in above-mentioned steps (B), add the copper ion inhibitor of 0.001%, finally add the Zulkovsky starch of 8%.
Again on the one hand, that the present invention's protection is described genetically engineered bacteria strain trichoderma asperellum (Tricho
derma asperellum) GY20 and the application of above-mentioned arbitrary described paste biological prevention and control agent in control cucumber fusarium axysporum.
According to following examples, can better understand the present invention.Concrete material proportion described in case study on implementation, processing condition and result thereof only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
Embodiment 1 paste biological prevention and control agent and preparation method thereof
The component of paste biological prevention and control agent is:
Active ingredient [trichoderma asperellum (
trichoderma asperellum) GY20 mycelia and chlamydospore mixture] 94.999%
Copper ion inhibitor MDA-5 0.001%
Potato starch 5%
PH value 3
Per-cent described above is weight percentage.
The preparation method of above-mentioned paste biological prevention and control agent comprises the following steps:
(A) acquisition of bacterium liquid
With aseptic inoculation cutter picking in advance at the upper cultured GY20 spore of potato dextrose agar (PDA), saving into spore content with aseptic washing downward is 10
5the inoculation liquid of spore/mL, by 5%(V/V) to be inoculated in the volume that 200mL potato grape liquid nutrient medium (PD) is housed be in the triangular flask of 750 mL to inoculum size, in 26 DEG C, shaking culture 48 h under 150 rpm/min conditions, it is in the triangular flask of 2000 mL that upper step gained nutrient solution 200 mL equal portions are inoculated in 4 volumes that 500mL PD nutrient solution is housed, shaking culture 48 h under 150 rpm, 26 DEG C of conditions; The 2000mL nutrient solution of gained is inoculated in the fermentor tank (east, Zhenjiang GUS-30) that the volume that 20L nutrient solution is housed is 30L, 20L fermentation culture liquid hold-up is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g, arranging fermentation condition is: dissolved oxygen 100%, stirring velocity is 350 rpm, leavening temperature 26 DEG C, fermentation time 48 h, pH 7.0-7.2.
(B) paste biological prevention and control agent complete processing
The fermented liquid fermented is processed into paste biological prevention and control agent as follows: first by fermented liquid centrifugal 10min under 8000rpm/min condition, obtain mycelia and chlamydospore mixture, with salt acid for adjusting pH to 3.0, and add the copper ion inhibitor of 0.001%, finally add the Zulkovsky starch of 5%, stir and namely made paste biological prevention and control agent.
Embodiment 2-3
Prepare embodiment 2-3 according to the method for embodiment 1, difference is as shown in table 1:
Table 1 embodiment 2-3 preparation method
| Paste biological prevention and control agent component/embodiment | Embodiment 2 | Embodiment 3 |
| Active ingredient | 89.95% | 91.99% |
| Copper ion inhibitor MDA-5 | 0.05% | 0.01% |
| Potato starch | 10% | 8% |
| PH value | 4.0 | 3.5 |
EXPERIMENTAL EXAMPLE 1:
The land for growing field crops application process of paste biological prevention and control agent of the present invention is as follows:
The application method of control cucumber fusarium axysporum is, by the 100L water dilution of 1Kg preparation, is first mixed with water by preparation, soaks root (soak 2 h) to use when cucumber seedling is transplanted.
Implementation result of the present invention
To cucumber fusarium axysporum field efficiency test
Experimental field condition: this test arrangement, in cucumber booth field, Vegetable Research room, Jiangsu Hilly Ground Zhenjiang Agriculture Science Research Institute, is continuous rich No. 2 for examination cucumber variety.
Test treatment process: dip in root process when cucumber seedling is transplanted.Namely, after the finished product GY20 paste biological prevention and control agent of embodiment 1 ~ 3 being pressed the dilution of 1Kg preparation 100L water, root process 2h is soaked when cucumber seedling is transplanted, blank clear water replaces, contrast biological and ecological methods to prevent plant disease, pests, and erosion medicament is " viride " wettable powder (production of Shandong Lunan biotechnology You Ji company limited), by 1Kg preparation with after the dilution of 100L water, root process 2h is soaked when cucumber seedling is transplanted, often process 300 seedlings, 3 are divided to repeat, community random alignment, transplants 50 d " Invest, Then Investigate " incidences, calculates diseased plant rate and preventive effect.
Result: greenhouse test result (table 2) shows, the prevention effect of GY20 to cucumber fusarium axysporum is all greater than more than 90%, and being significantly higher than " viride " to the prevention effect of cucumber fusarium axysporum is 58.4%.
Table 2 GY20 preparation is to the field controling test result of cucumber fusarium axysporum
Note: different capitalization represents the significance level of difference
p=0.01.
EXPERIMENTAL EXAMPLE 2:
The land for growing field crops application process of paste biological prevention and control agent of the present invention is as follows:
The application method of control cucumber fusarium axysporum is, by the 200L water dilution of 1Kg preparation, is first mixed with water by preparation, fills with root (every strain 200 mL) use when cucumber seedling is transplanted.
Implementation result of the present invention
To cucumber fusarium axysporum field efficiency test
Experimental field condition: this test arrangement, in cucumber booth field, Vegetable Research room, Jiangsu Hilly Ground Zhenjiang Agriculture Science Research Institute, is continuous rich No. 2 for examination cucumber variety.
Test treatment process: root irrigation when cucumber seedling is transplanted.Namely, after the finished product GY20 paste biological prevention and control agent of embodiment 1 ~ 3 being pressed the dilution of 1Kg preparation 200L water, fill with root (every strain 200 mL) when cucumber seedling is transplanted, blank clear water replaces, contrast biological and ecological methods to prevent plant disease, pests, and erosion medicament is (production of Shandong Lunan biotechnology You Ji company limited), by 1Kg preparation with after the dilution of 200L water, fill with root (every strain 200 mL) when cucumber seedling is transplanted, often process 300 seedlings, 3 are divided to repeat, community random alignment, transplants 50 d " Invest, Then Investigate " incidences, calculates diseased plant rate and preventive effect.
Result: greenhouse test result (table 3) shows, the prevention effect of GY20 to cucumber fusarium axysporum is all greater than more than 90%, and being significantly higher than " viride " to the prevention effect of cucumber fusarium axysporum is 57.24%.
Table 3 GY20 preparation is to the field controling test result of cucumber fusarium axysporum
Note: different capitalization represents the significance level of difference
p=0.01.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. a paste biological prevention and control agent for cucumber fusarium axysporum, is characterized in that, the activeconstituents of described preparation be trichoderma asperellum (
trichoderma asperellum) mycelia of GY20 and chlamydospore; The deposit number of described trichoderma asperellum GY20 is: CGMCC NO:4899;
Described mycelia and chlamydosporic ratio, determined by following cultural method:
By the trichoderma asperellum GY20 spore grown on potato dextrose agar with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, is inoculated in potato dextrose broth by 5% (V/V) inoculum size, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato dextrose broth shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain;
The weight part content of each component of the fermentation culture in described fermentor tank is: beet liquid 380-420, magnesium sulfate heptahydrate 3-5, potassium primary phosphate 38-42; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 300-400 rev/min, leavening temperature 25-27 degree Celsius, fermentation time 48 hours, pH 7.0-7.2; The condition of described shaking culture is 140-160 rev/min, 25-27 degree Celsius; Condition centrifugal after described fermentation culture is 7500-8500 rev/min, centrifugal 10 minutes.
2. the paste biological prevention and control agent of cucumber fusarium axysporum according to claim 1, is characterized in that, the fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2; The condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Condition centrifugal after described fermentation culture is 8000 revs/min, centrifugal 10 minutes;
Described paste biological prevention and control agent also comprises the copper ion inhibitor of weight percent 0.001 ~ 0.05%.
3. the paste biological prevention and control agent of cucumber fusarium axysporum according to claim 2, is characterized in that, described paste biological prevention and control agent also comprises the Zulkovsky starch of weight percent 5 ~ 10%;
Described copper ion inhibitor is MDA-5.
4. the paste biological prevention and control agent of cucumber fusarium axysporum according to claim 3, is characterized in that, described Zulkovsky starch is potato starch.
5. prepare a method for the paste biological prevention and control agent of cucumber fusarium axysporum according to claim 1, it is characterized in that, described method comprises:
1) by the trichoderma asperellum GY20 spore grown on potato dextrose agar with under aseptic washing, being adjusted to spore content is 10
5the inoculation liquid of spore/mL, by 5%(V/V) inoculum size is inoculated in potato dextrose broth, shaking culture 48 hours; By upper step gained nutrient solution with 10%(V/V) inoculum size to be again inoculated in potato dextrose broth shaking culture 48 hours further after amplification, then by gained nutrient solution with 10%(V/V) inoculum size be again inoculated in containing 20L fermentation culture fermentor tank in carry out fermentation culture after centrifugally to obtain;
2) adjust ph: the pH value of mycelia and chlamydospore mixture is adjusted to 3.0 ~ 4.0;
3) obtained paste biological prevention and control agent: to described step 2) add in the mixture that obtains 0.001 ~ 0.05% copper ion inhibitor, finally add the Zulkovsky starch of 5 ~ 10%, stir.
6. the method preparing the paste biological prevention and control agent of cucumber fusarium axysporum according to claim 5, is characterized in that, the condition of described shaking culture is 150 revs/min, 26 degrees Celsius; Fermentation culture liquid hold-up in described fermentor tank is: beet liquid 400.0g, magnesium sulfate heptahydrate 4.0g, potassium primary phosphate 40.0g; The condition of described fermentation is: dissolved oxygen 100%, and stirring velocity is 350 revs/min, leavening temperature 26 degrees Celsius, fermentation time 48 hours, pH 7.0-7.2; Condition centrifugal after described fermentation culture is 8000 revs/min, centrifugal 10 minutes; Described Zulkovsky starch is potato starch.
7. the application of paste biological prevention and control agent in control cucumber fusarium axysporum of cucumber fusarium axysporum according to claim 1.
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| CN103013838B (en) * | 2012-12-14 | 2015-05-13 | 江苏耕耘化学有限公司 | Pasty biocontrol preparation for strawberry greensickness, and preparation method, application and special strain thereof |
| CN103184162B (en) * | 2013-04-24 | 2014-11-19 | 牛赡光 | Trichoderma asperellum and applications thereof |
| CN103289902B (en) * | 2013-05-02 | 2014-12-10 | 华南农业大学 | Trichoderma asperellum and biological preparation and application in controlling and preventing cucumber phytophthora root rot |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057593A (en) * | 2007-01-26 | 2007-10-24 | 中国计量学院 | Preparation method for preventing and treating vegetable fungicide microbial pesticides |
| CN101874484A (en) * | 2009-12-18 | 2010-11-03 | 江苏耕耘化学有限公司 | Paste dosage-form trichoderma biopesticide composite and preparation method thereof |
| CN102067885A (en) * | 2010-03-31 | 2011-05-25 | 惠州市南天生物科技有限公司 | Trichoderma spore powder as well as preparation method and application thereof |
| CN103013838A (en) * | 2012-12-14 | 2013-04-03 | 江苏耕耘化学有限公司 | Pasty biocontrol preparation for strawberry greensickness, and preparation method, application and special strain thereof |
-
2012
- 2012-12-14 CN CN201210542063.0A patent/CN103013839B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057593A (en) * | 2007-01-26 | 2007-10-24 | 中国计量学院 | Preparation method for preventing and treating vegetable fungicide microbial pesticides |
| CN101874484A (en) * | 2009-12-18 | 2010-11-03 | 江苏耕耘化学有限公司 | Paste dosage-form trichoderma biopesticide composite and preparation method thereof |
| CN102067885A (en) * | 2010-03-31 | 2011-05-25 | 惠州市南天生物科技有限公司 | Trichoderma spore powder as well as preparation method and application thereof |
| CN103013838A (en) * | 2012-12-14 | 2013-04-03 | 江苏耕耘化学有限公司 | Pasty biocontrol preparation for strawberry greensickness, and preparation method, application and special strain thereof |
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