CN103805527A - StreptomycesYunnanensis var daliensis and fermentation broth and application thereof - Google Patents
StreptomycesYunnanensis var daliensis and fermentation broth and application thereof Download PDFInfo
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- CN103805527A CN103805527A CN201210438796.XA CN201210438796A CN103805527A CN 103805527 A CN103805527 A CN 103805527A CN 201210438796 A CN201210438796 A CN 201210438796A CN 103805527 A CN103805527 A CN 103805527A
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Abstract
The invention discloses a StreptomycesYunnanensis var daliensis and a fermentation broth thereof. The 16Sr DNA sequence of the strain is shown as a SEQ ID NO.1; the strain has a preservation number of CGMCC No.5912, can be used for control of plant viral diseases, including tobacco mosaic virus, pepper virus disease, rice black-streaked dwarf virus and cabbage brown streak disease, and has wide application prospects.
Description
Technical field
The present invention relates to microbial technology field, specifically, relate to the Dali mutation of a kind of Yunnan streptin and fermentation culture and application.
Background technology
The diversity of ecotope has brought up the specificity of the various and pathways metabolism of microbe species.Conventionally, can be by the morphological specificity of microorganism strains, cultural characteristic and physiological and biochemical property etc. to the microbial strains discriminating of classifying.Along with the development of Protocols in Molecular Biology, the normal classification position that carrys out clear and definite bacterial strain in conjunction with the conserved dna sequence that detects microorganism strains at present.Due to microorganism, to have growth cycle short, and metabolism is easy to regulation and control, can realize by fermentation the feature of scale operation, so microorganism is the Main Resources of biological pesticide always.
Virus disease is intractable disease serious to murrain and that be difficult to control, once crop infects virus disease, will there will be floral leaf, leaflet, tender leaf or plant to occur streak, the rugged shape of fruit, and even complete stool is withered.China's common Disease in field mainly contains at present: stripe disease, black streaked dwarf virus of rice, tobacco mosaic disease, vegetables mosaic virus, potato mosaic viral disease, tomato virus disease pepper virus disease etc.The biological pesticide for preventing and treating Disease that China registers on the books at present only has one.
Summary of the invention
One of object of the present invention is to provide a kind of Yunnan streptin Dali mutation (StreptomycesYunnanensis var daliensis), the 16Sr DNA sequence dna of this bacterial strain is as shown in SEQ ID NO.1, be deposited in Chinese common micro-organisms preservation center, preserving number is CGMCC No.5912.
Two of object of the present invention is to provide this Yunnan streptin Dali mutation (StreptomycesYunnanensis var daliensis) for preventing and treating the application of plant virus venereal disease evil.
Wherein, plant virus venereal disease evil comprises: tobacco mosaic virus (TMV), capsicum virus, black streaked dwarf virus of rice, Chinese cabbage brown streak.
It is a kind of for preventing and treating the fermentation culture of plant virus venereal disease evil that three of object of the present invention is to provide, and this fermentation culture is the fermentation culture of described Yunnan streptin Dali mutation (StreptomycesYunnanensis var daliensis).
The culture condition of described fermentation culture is: culture temperature is 25-30 ℃, and incubation time is 72-120 hour; Culture medium prescription comprises: groundnut meal, starch, soluble starch, Semen Maydis powder, glucose, yeast powder, salt, calcium carbonate, ammonium sulfate, potassium primary phosphate, magnesium sulfate.
Yunnan streptin disclosed by the invention Dali mutation (Streptomyces Yunnanensis vardaliensis) is that the method by adopting native separate microorganism obtains, its fermentation culture can be used as biological pesticide, comprise the plant virus venereal disease evils such as tobacco mosaic virus (TMV), capsicum virus, black streaked dwarf virus of rice, Chinese cabbage brown streak for control, there is prospect of the application widely.Because the microbial growth cycle is short, metabolism is easy to regulation and control, the therefore very applicable scale operation of fermentation technique.
Accompanying drawing explanation
Fig. 1 is the electromicroscopic photograph of the mycelia form of Yunnan streptin Dali mutation (Streptomyces Yunnanensis var daliensis).
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.Should be understood that following examples are only for the present invention is described but not for limiting scope of the present invention.
Yunnan streptin of the present invention Dali mutation (S.Yunnanensis var daliensis) is that the method by adopting native separate microorganism obtains, its bacterial strain on March 15th, 2012 in Chinese common micro-organisms preservation center preservation (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preserving number: CGMCC No.5912.Accompanying drawing 1 obtains viability report for microbial preservation.
(1), the physicochemical property of bacterial strain
1, morphological specificity and cultural characteristic
The fibrillae of spores of Yunnan streptin Dali mutation (S.Yunnanensis var daliensis) bacterial strain is flexible, hook-type, and spore column, ellipse, refer to accompanying drawing 1.The growth characteristics of bacterial strain on various substratum is as shown in table 1.
Table 1
2, Physiology and biochemistry and utilization of carbon source feature
Physiology and biochemistry and the utilization of carbon source feature of Yunnan streptin Dali mutation (S.Yunnanensis var daliensis) bacterial strain are as shown in table 2.
Table 2
3, DNA sequence dna
The 16SrDNA sequence of Yunnan streptin Dali mutation (S.Yunnanensis var daliensis) bacterial strain is as shown in SEQ ID NO.1.
Above-mentioned form, physiological and biochemical property and 16S rDNA sequential analysis show, this bacterial strain belongs to Yunnan streptin (Yunnanensis) the Dali mutation (vardaliensis) of streptomyces (Streptomyces), therefore name as Yunnan streptin (Streptomyces Yunnanensis var daliensis) DL-101.
The cultural method of this bacterial strain both can be cultivated for deep-layer liquid, can be also solid culture.For being conducive to payable cultivation, the spore suspension of above-mentioned production bacterium should be inoculated on substratum to the gas agitating cultivation that works of going forward side by side.
(2), embodiment 1: the cultivation of bacterial strain
Formula according to embodiment 1 is made substratum, pH regulator to 7.0, sterilization.Yunnan streptin Dali mutation (S.Yunnanensis var daliensis) the DL-101 bacterial classification that grows in oatmeal medium slant is inoculated in this substratum, at 28 ℃, cultivates 30 hours.Take this culture as seed, be inoculated in the fermentation culture of above-mentioned substratum composition with 10% ratio, at 28 ℃, aeration-agitation is cultivated 100 hours again.After fermentation culture finishes, fermented liquid can be directly used in controlling plant diseases, also simply after concentration, applies again.
The culture medium prescription of embodiment 1: groundnut meal 3%, starch 1%, soluble starch 1.5%, Semen Maydis powder 1%, glucose 0.5%, yeast powder 0.1%, salt 0.3%, calcium carbonate 0.3%, ammonium sulfate 0.25%, potassium primary phosphate 0.025%, magnesium sulfate 0.0025%.
Nutrition source in culture medium prescription there is no special requirement, can make to contain in substratum the carbon source, nitrogenous source and other nutrition sources that are usually used in microorganism culturing.
Wherein, carbon source can be in aforementioned utilization of carbon source table arbitrarily available one or more; Nitrogenous source can be other organic or inorganic nitrogenous compounds such as meat peptone, soybean cake powder, groundnut meal, yeast, meat extract, corn steep liquor, cottonseed meal, urea, ammonium salt, nitrate; Other nutrition sources can add some inorganic salts, and the metal-salts such as such as salt, phosphoric acid salt and potassium, calcium, magnesium, iron, manganese, also can add some animals and plants, mineral wet goods if desired as defoamer.
The conditions such as the temperature of the fermentation of this bacterial strain to substratum, pH, time there is no strict restriction.Active ingredient in fermented liquid is higher, is more conducive to the production of bacterial strain, and test shows, under the following conditions, active ingredient output is higher: glucose accounts for 3%, starch accounts for 2% left and right; The pH scope of substratum is 6-8, neutral for well to approach; Culture temperature is at 20-35 ℃; Ventilation ratio is (ventilation ratio: per minute is by the volume of air of nutrient solution and the ratio of nutrient solution volume) between 1:0.2-1:3.The component of above-mentioned substratum, pH, culture temperature, aeration condition etc. also should suitably be adjusted according to practical situation, to obtain best effect.
(3) application of bacterial strain
Yunnan streptin Dali mutation (S.Yunnanensis var daliensis) DL-101 mainly can be used for preventing and treating various plants Disease, comprises the disease that the multiple viruses such as tobacco mosaic virus (TMV), capsicum virus, black streaked dwarf virus of rice, Chinese cabbage brown streak cause.
Embodiment 2: for the preventive effect of tobacco mosaic virus (TMV)
By the fermentation culture dilute with water 20-50 making according to embodiment 1 doubly, then even spraying is on the 4-6 of the indoor cultivation tobacco of leaf phase; Within second day, adopt the mode of artificial frictional inoculation tobacco mosaic virus (TMV) to contaminate; After one day, medication is once again to connect poison; After 7 days, medication once, shares medicine three times again.
Preventive effect is as shown in table 3.
Table 3
In table, disease index is the overall target of comprehensively considering sickness rate and severity.Calculation formula is: in the time that severity represents by classification typical value, and disease index=100 × ∑ (the sick numbers of sheets at different levels × typical values at different levels)/(investigating total number of sheets × highest typical value); In the time that severity represents with percentage, disease index=generally rate × severity.
Relative control effect refers to chemicals treatment and the blank prevention effect after relatively, and calculation formula is: relative control effect=(disease index of blank disease index-chemicals treatment)/blank disease index × 100%.
Embodiment 3: for the result for the treatment of of tobacco mosaic virus (TMV), capsicum virus, Chinese cabbage brown streak
In field, be chosen in tobacco, capsicum, the Chinese cabbage of their early stage, by the culture making according to embodiment 1, even spraying after 20 ~ 50 times of dilute with waters, every medication in 7 days once, continuous use 3 times.For the treatment result of tobacco mosaic virus (TMV), capsicum virus, Chinese cabbage brown streak respectively as shown in table 4, table 5, table 6.
Table 4
Table 5
Table 6
Embodiment 4, for the integrated control effect of black streaked dwarf virus of rice
In field, be chosen in the field that planthopper has just migrated into, by after the pymetrozine WDG 100g mixture of the culture 1kg making according to embodiment 1 and 25%, 50kg adds water.After stirring, spraying is processed, and every medication in 7 days once, then detects its prevention effect to black streaked dwarf virus of rice.Result demonstration, its integrated control effect to black streaked dwarf virus of rice reaches 95%.
Claims (6)
1. a Yunnan streptin Dali mutation (Streptomyces Yunnanensis vardaliensis), it is characterized in that: the 16Sr DNA sequence dna of this bacterial strain is as shown in SEQ ID NO.1, be deposited in Chinese common micro-organisms preservation center, preserving number is CGMCC No.5912.
2. Yunnan streptin as claimed in claim 1 Dali mutation (S.Yunnanensis vardaliensis) is for preventing and treating the application of plant virus venereal disease evil.
3. application according to claim 2, is characterized in that: described plant virus venereal disease evil comprises: tobacco mosaic virus (TMV), capsicum virus, black streaked dwarf virus of rice, Chinese cabbage brown streak.
4. for preventing and treating a fermentation culture for plant virus venereal disease evil, it is characterized in that: the fermentation culture that is Yunnan streptin as claimed in claim 1 Dali mutation (S.Yunnanensis vardaliensis).
5. fermentation culture according to claim 4, is characterized in that: the culture condition of described fermentation culture is: culture temperature is 25-30 ℃, and incubation time is 72-120 hour.
6. fermentation culture according to claim 4, is characterized in that: the culture medium prescription of described fermentation culture comprises: groundnut meal, starch, soluble starch, Semen Maydis powder, glucose, yeast powder, salt, calcium carbonate, ammonium sulfate, potassium primary phosphate, magnesium sulfate.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106305726A (en) * | 2016-08-23 | 2017-01-11 | 乳山韩威生物科技有限公司 | Ectoines wettable powder containing brassinolide as well as preparation method and application thereof |
| CN110408559A (en) * | 2019-07-05 | 2019-11-05 | 青岛农业大学 | One plant of rose streptomyces verticillus and its application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1358841A (en) * | 2000-12-11 | 2002-07-17 | 云南省微生物研究所 | Yunnan streptin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1358841A (en) * | 2000-12-11 | 2002-07-17 | 云南省微生物研究所 | Yunnan streptin |
Non-Patent Citations (2)
| Title |
|---|
| QI ZHANG ET AL.: "Streptomyces yunnanensis sp. nov., a mesophile from soils in Yunnan, China", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 * |
| 谭仲夏: "云南链霉菌的发酵及其产物放线酮对烟草病害的防治研究", 《中国优秀硕士学位论文全文数据库(电子期刊)农业科技辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106305726A (en) * | 2016-08-23 | 2017-01-11 | 乳山韩威生物科技有限公司 | Ectoines wettable powder containing brassinolide as well as preparation method and application thereof |
| CN106305726B (en) * | 2016-08-23 | 2018-10-30 | 乳山韩威生物科技有限公司 | A kind of pyrimidine acid rhzomorph wettable powder and its preparation method and application containing brassin lactones |
| CN110408559A (en) * | 2019-07-05 | 2019-11-05 | 青岛农业大学 | One plant of rose streptomyces verticillus and its application |
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