CN104388356A - Streptomyces sampsonii strain as well as separation method and application thereof - Google Patents
Streptomyces sampsonii strain as well as separation method and application thereof Download PDFInfo
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- CN104388356A CN104388356A CN201410705973.5A CN201410705973A CN104388356A CN 104388356 A CN104388356 A CN 104388356A CN 201410705973 A CN201410705973 A CN 201410705973A CN 104388356 A CN104388356 A CN 104388356A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention relates to the technical field of microbial pesticides, and particularly provides a streptomyces sampsonii strain, a separation method and a culture method of the streptomyces sampsonii strain, a streptomyces sampsonii culture obtained by using the culture method, applications of the streptomyces sampsonii strain and the streptomyces sampsonii culture, and a pesticide containing the streptomyces sampsonii strain and the streptomyces sampsonii culture. The preservation name of the streptomyces sampsonii strain is streptomyces sampsonii (Streptomyces sampsonii) SY-FX-31; the streptomyces sampsonii is preserved at the China Center for Type Culture Collection; the preservation number is CCTCC M2014516; and a gene sequence of the strain 16S rRNA is as shown in a sequence table SEQ ID No.1. The streptomyces sampsonii strain is capable of preventing and treating a plurality of plant diseases, is high in growth speed, large in sporulation quantity and wide in antimicrobial spectrum, and can be quickly colonized in soil and plant bodies.
Description
Technical field
The present invention relates to microbial pesticide technical field, be specifically related to a kind of Sang Puxun streptomycete bacterial strain and separation method thereof and application.
Background technology
Microbial pesticide is due to its environmental protection, and the characteristic not easily developed immunity to drugs, obtains paying attention to more and more widely and applying in controlling crop diseases and insect pests.The promotion and application technology of microbial pesticide is tending towards ripe just gradually, country strengthens year by year in the policy support dynamics of field of biological control, the use of microbial pesticide can overcome chemical pesticide to the pollution of ecotope and to reduce at agricultural byproducts Pesticides residual, promote quality and the price of agricultural byproducts, not only there is environmental benefit but also there is economic benefit.
At present, major microorganisms pesticide species market promoted concentrates on bacillus and Rhodopseudomonas.Streptomyces is that a class can produce antibiotic microorganism Pseudomonas, be Streptomyces griseoviridis and Streptomyces lydicus as agricultural fungicidal class in the kind that US and European is registered, streptomyces is expected to become the another kind of microbial pesticide in field of biological control widespread use after bacillus and Rhodopseudomonas.
Summary of the invention
The invention provides a kind of newly can prevent and treat plurality of plant diseases, and fast growth, sporulation quantity are large, antimicrobial spectrum is wide, surely the Sang Puxun streptomycete bacterial strain grown rapidly in soil and plant physical efficiency and separation method thereof and cultural method, and by the Sang Puxun Streptomyces culture of this cultural method gained, and the application of this Sang Puxun streptomycete bacterial strain and Sang Puxun Streptomyces culture and containing their agricultural chemicals.
A first aspect of the present invention provides a kind of Sang Puxun streptomycete bacterial strain (Streptomycessampsonii) SY-FX-31, preservation name is called Sang Puxun streptomycete SY-FX-31 (Streptomycessampsonii), depositary institution: China typical culture collection center, depositary institution address: Wuhan University's Life Science College, preservation date is: on October 27th, 2014, deposit number is: CCTCCM 2014516, and this bacterial strain 16S rRNA gene order is as shown in sequence table SEQ ID No.1.
This bacterial strain has following biological characteristics: (1), in the growth of Gause I substratum, mycelia is light yellow, and along with incubation time extends, mycelia darkens.(2) examining the growth of protein peptone substratum, mycelia khaki color, there is orange pigment in bacterium colony and substratum junction.(3) light yellow maize powder medium growth mycelia, after gradually become deep yellow, in light yellow hypha body in liquid culture.
Its 16S rRNA gene order sequencing result shows that the 16S rRNA gene order of SY-FX-31 bacterial strain is the nucleotide sequence in sequence table SEQ ID No.1.Carry out tetraploid rice according to sequencing result and Genbank Streptomyces 16S rRNA gene order, result shows the Sang Puxun streptomycete of SY-FX-31 Pseudomonas in streptomyces (Streptomyces) (Streptomyces sampsonii).
A second aspect of the present invention is provided for the separation method being separated above-mentioned Sang Puxun streptomycete bacterial strain, and its step comprises: S1, gathers earth from river bed, earth is mixed with sterilized water scatter after adding sterilized water vibration, staticly makes it clarify, and gets supernatant liquor; S2, adopt plate streak separation and purification Sang Puxun streptomycete bacterial strain: diluted by step S1 gained supernatant liquor, the diluent of different weaker concn is adopted to coat on Gause I culture medium flat plate after 10-40 DEG C of cultivation 2-7d, picking list bacterium colony separation and purification get Sang Puxun streptomycete bacterial strain; Wherein, the composition of Gause I substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6.
Wherein, plate streak refers to and the different cell transfering loops in microorganism mixed in together or same microbial population is diluted by sectional streak at slat chain conveyor primary surface and obtains the individual cells of more independent distribution, after cultivating, growth and breeding becomes single bacterium colony, usually this single bacterium colony is used as the purebred of microorganism to be separated.Sometimes this single bacterium colony is not all bred by individual cells, can adopt repeatedly be separated repeatedly obtain purebred.Microbiological specimens is repeatedly done " by point to line " dilution at solid culture primary surface and reaches separation object by its principle.The form of line has multiple, rule in the community be specifically as follows a flat board is divided into four different area, firstth district (A district) area is minimum, as the bacterium source region of bacterium to be separated, second and the 3rd district (B, C district) be the zone of transition of stepwise dilution, 4th district (D district) is then key area, and a large amount of single bacterium colonies purebred use for you to choose appears in Shi Gai district.In order to obtain the single bacterium colony of more typical case, the distribution of dull and stereotyped upper four district's areas should be D > C > B > A.
A third aspect of the present invention provides the cultural method of above-mentioned Sang Puxun streptomycete bacterial strain, and its step comprises S1, by above-mentioned Sang Puxun streptomycete bacterial strain in storage medium, at 25-33 DEG C, activates 12-24h; S2, by the bacterial strain after activation in seed culture medium, cultivates 1-2 days in 25-33 DEG C, 100-150r/min, obtains seed liquor; S3, is inoculated in fermention medium by gained seed liquor with volumn concentration 4-8% inoculative proportion, and at 25-33 DEG C, 100-150r/min shaking culture 2-4 days obtains fermented liquid; Wherein, preservation isolation medium is Gause I substratum, and the composition of described Gause I substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6; Seed culture medium for examining protein peptone substratum, described in examine protein peptone substratum composition comprise: sucrose, peptone, FeSO
4, MgSO
4, KCl, K
2hPO
4and water; Fermention medium is maize powder medium, and the composition of described maize powder medium comprises: Semen Maydis powder, sucrose, peptone, starch, yeast extract paste, NaCl, K
2hPO
4, MgSO
4, CaCO
3and water.
A fourth aspect of the present invention provides a kind of Sang Puxun Streptomyces culture, is obtained by above-mentioned cultural method.
A fifth aspect of the present invention provides above-mentioned Sang Puxun streptomycete bacterial strain and/or the application of its culture in controlling plant diseases.
A sixth aspect of the present invention provides a kind of agricultural chemicals, and this agricultural chemicals comprises above-mentioned Sang Puxun streptomycete bacterial strain and/or its culture.
The Sang Puxun streptomycete (Streptomyces sampsonii) of gained of the present invention belongs to streptomyces, finds that it is to gibberella saubinetii through indoor biological determination of activity, and rice banded sclerotial blight etc. tens kind of plant pathogenic bacteria all has restraining effect.And the Sang Puxun streptomycete fast growth found, sporulation quantity is large, antimicrobial spectrum is wide, surely grow rapidly in soil and plant physical efficiency.Can not only prevent and treat plurality of plant diseases, and can improve plant resistance to environment stress, all having active effect to raising crop yield and lifting quality, is a kind of biocontrol strains having application prospect.
Embodiment
In order to make technical problem solved by the invention, technical scheme and beneficial effect clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
The invention provides a kind of Sang Puxun streptomycete bacterial strain, preservation name is called Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31, be preserved in China typical culture collection center, deposit number is: CCTCC M 2014516, this bacterial strain 16S rRNA gene order is as shown in sequence table SEQ IDNo.1, belong to streptomyces, plurality of plant diseases can not only be prevented and treated, and can plant resistance to environment stress be improved, all having active effect to raising crop yield and lifting quality, is a kind of biocontrol strains having application prospect.
Invention also provides the separation method for separating of above-mentioned Sang Puxun streptomycete bacterial strain, its step comprises: S1, earth is gathered from river bed, after adding sterilized water vibration, earth is mixed to scatter with sterilized water, be specifically as follows in container earth being added in-built sterilized water and granulated glass sphere, for some time of vibrating on shaking table makes earth scatter, and vibration terminates staticly to make it clarify, and gets supernatant liquor.S2, adopts plate streak separation and purification Sang Puxun streptomycete bacterial strain: be specially and diluted by step S1 gained supernatant liquor, and concrete dilution process can adopt and add the dilution of sterilized water vortex mixed; Adopt the diluent of different weaker concn to coat on Gause I culture medium flat plate, be specifically as follows each concentration be coated with 3 parallel, cultivate after 2-7d in 10-40 DEG C, picking list bacterium colony separation and purification get Sang Puxun streptomycete bacterial strain.Wherein, the composition of Gause I substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6.
Wherein, Sang Puxun streptomycete bacterial strain is in the growth of Gause I substratum, and mycelia is light yellow, and along with incubation time extends, mycelia darkens.
Preferably, the composition of Gause I substratum comprises: Zulkovsky starch 5-40g, KNO
30.1-5g, NaCl 0.1-3g, K
2hPO
43H
2o 0.1-3g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-3g and water 1-10L.
Preferably, dilute three concentration, the concentration of diluent comprises supernatant liquor dilution 10
-2, 10
-3, 10
-4doubly, be spread evenly across respectively on Gause I culture medium flat plate.
Invention also provides the cultural method of above-mentioned Sang Puxun streptomycete bacterial strain, its step comprises S1, by above-mentioned Sang Puxun streptomycete bacterial strain in storage medium, at 25-33 DEG C, activates 12-24h; S2, by the bacterial strain after activation in seed culture medium, cultivates 1-2 days in 25-33 DEG C, 100-150r/min, obtains seed liquor; S3, is inoculated in fermention medium by gained seed liquor with 4-8% (v/v) inoculative proportion, and at 25-33 DEG C, 100-150r/min shaking culture 2-4 days obtains fermented liquid.
Wherein, storage medium is Gause I substratum, and the composition of described Gause I substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6.Preferably concrete, the composition of Gause I substratum comprises: Zulkovsky starch 5-40g, KNO
30.1-5g, NaCl 0.1-3g, K
2hPO
43H
2o 0.1-3g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-3g and water 1-10L.
Wherein, seed culture medium for examining protein peptone substratum, described in examine protein peptone substratum composition comprise: sucrose, peptone, FeSO
4, MgSO
4, KCl, K
2hPO
4and water; Preferably, the composition examining protein peptone substratum comprises: sucrose 5-50g, peptone 1-15g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-5g, KCl 0.1-5g, K
2hPO
40.1-5g and water 1-10L.
Wherein, fermention medium is maize powder medium, and the composition of described maize powder medium comprises: Semen Maydis powder, sucrose, peptone, starch, yeast extract paste, NaCl, K
2hPO
4, MgSO
4, CaCO
3and water; Preferably, the composition of maize powder medium comprises: Semen Maydis powder 5-50g, sucrose 1-20g, peptone 1-10g, starch 1-20g, yeast extract paste 1-10g, NaCl 1-20g, K
2hPO
40.1-5g, MgSO
40.1-5g, CaCO
30.1-5g and water 1-10L.
Invention also provides a kind of Sang Puxun Streptomyces culture, obtained by above-mentioned cultural method.
Invention also provides the application of above-mentioned Sang Puxun streptomycete bacterial strain and/or its culture, this Sang Puxun streptomycete bacterial strain and/or its culture are used for controlling plant diseases.
Wherein, Plant diseases comprises that white muskmelon melon is rotten, capsicum withers, tomato early epidemic, cucumber anthrax, cotton yellow wither, cucumber grey mold, rice bakanae disease, tomato green grass or young crops are withered, one or more in rice banded sclerotial blight, the black shin of tobacco, Root Rot of Wheat, paddy rice rice blast, gibberella saubinetii, rape sclerotium, soybean root rot, tomato gray mould, cucumber foxiness or cucumber downy mildew, multiple disease and pest can be prevented, antimicrobial spectrum is wide, and application prospect is wide.
Invention also provides a kind of agricultural chemicals, this agricultural chemicals comprises above-mentioned Sang Puxun streptomycete bacterial strain and/or its culture, is microbial pesticide, can obtain or carry out formulation and obtain for above-mentioned obtained fermented liquid directly dilutes.
Below in conjunction with specific embodiment, the present invention is further described.
embodiment 1
The separation andpreconcentration of Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31
1. the separation of Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31
Gather soil 10g from river bed, add in the triangular flask of in-built 100mL sterilized water and granulated glass sphere, vibrate 30min on the shaking table of 120r/min, and vibration terminates rear standing 1h, and get supernatant 0.5mL and add in 4.5mL sterilized water, vortex mixed, dilutes 10 successively
-2, 10
-3, 10
-4doubly, get 150 μ L diluents respectively and coat on Gause I culture medium flat plate, each concentration be coated with 3 parallel, cultivate 2-7d at 28 DEG C, the single bacterium colony on picking substratum carries out plate streaking purifying, obtains Sang Puxun streptomycete bacterial strain.
2. the qualification of Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31
16S rRNA gene order sequencing result shows that the 16S rRNA gene order of SY-FX-31 bacterial strain is sequence table SEQ ID No.1 nucleotide sequence.Carry out tetraploid rice according to sequencing result and Genbank Streptomyces 16S rRNA gene order, result shows the Sang Puxun streptomycete of SY-FX-31 Pseudomonas in streptomyces (Streptomyces) (Streptomyces sampsonii).
embodiment 2
Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31 tests pathogenic fungi isolated activity
By Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31 in storage medium (Zulkovsky starch 20g, KNO
31g, NaCl 0.5g, K
2hPO
43H
2o 0.5g, FeSO
47H
2o 0.01g, MgSO
47H
2o 0.5g, water 1L, pH7.4-7.6) in, activate 20h at 25 DEG C after, by the bacterial strain after activation in seed culture medium (sucrose 30g, peptone 5g, FeSO
47H
2o 0.01g, MgSO
47H
2o 0.5g, KCl 0.5g, K
2hPO
41g, water 1L) in, in 25 DEG C, 150r/min cultivates 2 days, obtains seed liquor; Then gained seed liquor is inoculated in fermention medium (Semen Maydis powder 20g, sucrose 10g, peptone 2g, starch 5g, yeast extract paste 2g, NaCl 2g, K with 5% (v/v) inoculative proportion
2hPO
40.5g, MgSO
40.5g, CaCO
31g, water 1L) in, at 25 DEG C, 150r/min shaking culture obtains fermented liquid in 2 days.
Fermented liquid is mixed with certain density mother liquor, adopt containing toxic medium method, PDA substratum (potato 200 grams, glucose 20 grams, 20 grams, agar, water 1L) in add the mother liquor prepared in advance, make pastille substratum, after cooling, aseptically inoculation supplies examination pathogenic bacteria, with adding the PDA substratum of sterilized water in contrast.Form bacterium colony for the sick fungi of examination in the dull and stereotyped activation culture of PDA, produce bacterium cake with punch tool (5mm), mycelia is faced down and connects bacterium in the dull and stereotyped central authorities of PDA.The flat board inoculated is placed in 28 DEG C of biochemical cultivation cases and cultivates 4-7 days " Invest, Then Investigate "s, adopt right-angled intersection method to measure colony growth diameter, and calculate bacteriostasis rate.
Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31 is as shown in table 1 to the restraining effect result for examination pathogenic fungi.
Table 1
Sang Puxun streptomycete of the present invention (Streptomyces sampsonii) SY-FX-31 is good to the restraining effect of pathogenic fungi as can be seen from the above table.
Embodiment 3
The living body biological determination of activity of Sang Puxun streptomycete (Streptomyces sampsonii) SY-FX-31
The potted plant cucumber seedling (1 leaf 1 heart stage) of growth selection neat and consistent, above-mentioned obtained fermented liquid is mixed with finite concentration on crops sprayer, carries out spraying process, clear water contrasts, and potted plant seedling is naturally dried after chemicals treatment.Inoculate bacterium of downy mildew of cucumber spore suspension after 24h and be placed in phytotron 24h (temperature: 23 DEG C, humidity: move to normal management in greenhouse 90%).Greenhouse experiment: temperature 22-30 DEG C, humidity 40-60%.
The pot rice seedling (3 leaf phase) of growth selection neat and consistent, above-mentioned obtained fermented liquid is mixed with finite concentration on crops sprayer, carries out spraying process, clear water contrasts, and potted plant seedling is naturally dried after chemicals treatment.After 24h bottom plant the withered mycelia block of Inoculated Rice line, be placed in phytotron 5-7 days (temperature: 28 DEG C, humidity: 80%), observation period incidences.
In greenhouse, cultivate 7d investigation prevention effect, grade scale performs National Standard of the People's Republic of China " pesticide field efficacy medicine test criterion () ", calculates prevention effect with disease index.Result is as shown in table 2.
Drug effect method of calculation
Table 2
Prevent and treat result as can be seen from live body test, the fermented liquid of Sang Puxun streptomycete (Streptomycessampsonii) SY-FX-31 of the present invention has certain prevention effect to rice banded sclerotial blight, cucumber downy mildew.
In the description of this specification sheets, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present invention or example.In this manual, to the schematic representation of above-mentioned term not must for be identical embodiment or example.And the specific features of description, structure, material or feature can combine in one or more embodiment in office or example in an appropriate manner.In addition, when not conflicting, the feature of the different embodiment described in this specification sheets or example and different embodiment or example can carry out combining and combining by those skilled in the art.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, and those of ordinary skill in the art can change above-described embodiment within the scope of the invention, revises, replace and modification.
Claims (10)
1. Yi Zhong Sang Puxun streptomycete bacterial strain, preservation name is called Sang Puxun streptomycete (Streptomycessampsonii) SY-FX-31, be preserved in China typical culture collection center, deposit number is: CCTCC M 2014516, and this bacterial strain 16S rRNA gene order is as shown in sequence table SEQ ID No.1.
2., for separating of a separation method for Sang Puxun streptomycete bacterial strain according to claim 1, it is characterized in that, comprise step:
S1, gathers earth from river bed, earth is mixed with sterilized water scatter after adding sterilized water vibration, staticly makes it clarify, and gets supernatant liquor;
S2, adopt plate streak separation and purification Sang Puxun streptomycete bacterial strain: diluted by step S1 gained supernatant liquor, after adopting the diluent of different weaker concn to coat Gause I culture medium flat plate cultivates 2-7d at 10-40 DEG C, picking list bacterium colony separation and purification get Sang Puxun streptomycete bacterial strain;
The composition of described Gause I substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6.
3. separation method according to claim 2, is characterized in that, described Sang Puxun streptomycete bacterial strain is in the growth of Gause I substratum, and mycelia is light yellow, and along with incubation time extends, mycelia darkens;
The composition of described Gause I substratum comprises: Zulkovsky starch 5-40g, KNO
30.1-5g, NaCl 0.1-3g, K
2hPO
43H
2o 0.1-3g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-3g and water 1-10L.
4. the separation method of Sang Puxun streptomycete bacterial strain according to claim 2, is characterized in that, the concentration of described diluent comprises supernatant liquor dilution 10
-2, 10
-3, 10
-4doubly.
5. a cultural method for Sang Puxun streptomycete bacterial strain as claimed in claim 1, is characterized in that, comprise step:
S1, by Sang Puxun streptomycete bacterial strain according to claim 1 in storage medium, activates 12-24h at 25-33 DEG C;
S2, by the bacterial strain after activation in seed culture medium, cultivates 1-2 days in 25-33 DEG C, 100-150r/min, obtains seed liquor;
S3, is inoculated in fermention medium by gained seed liquor with volumn concentration 4-8% inoculative proportion, and at 25-33 DEG C, 100-150r/min shaking culture 2-4 days obtains fermented liquid;
Described storage medium is Gause I substratum, and the composition of described Gao Shi No. 1 substratum comprises: Zulkovsky starch, KNO
3, NaCl, K
2hPO
4, FeSO
4, MgSO
4and water, pH value is 7.4-7.6;
Described seed culture medium for examining protein peptone substratum, described in examine protein peptone substratum composition comprise: sucrose, peptone, FeSO
4, MgSO
4, KCl, K
2hPO
4and water;
Described fermention medium is maize powder medium, and the composition of described maize powder medium comprises: Semen Maydis powder, sucrose, peptone, starch, yeast extract paste, NaCl, K
2hPO
4, MgSO
4, CaCO
3and water.
6. cultural method according to claim 5, is characterized in that,
The composition of described Gause I substratum comprises: Zulkovsky starch 5-40g, KNO
30.1-5g, NaCl 0.1-3g, K
2hPO
43H
2o 0.1-3g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-3g and water 1-10L;
The described composition examining protein peptone substratum comprises: sucrose 5-50g, peptone 1-15g, FeSO
47H
2o 0.01-1g, MgSO
47H
2o 0.1-5g, KCl 0.1-5g, K
2hPO
40.1-5g and water 1-10L;
The composition of described maize powder medium comprises: Semen Maydis powder 5-50g, sucrose 1-20g, peptone 1-10g, starch 1-20g, yeast extract paste 1-10g, NaCl 1-20g, K
2hPO
40.1-5g, MgSO
40.1-5g, CaCO
30.1-5g and water 1-10L.
7. Yi Zhong Sang Puxun Streptomyces culture, is obtained by the cultural method described in claim 5 or 6.
8. Sang Puxun streptomycete bacterial strain according to claim 1 and/or the application of Sang Puxun Streptomyces culture according to claim 7 in controlling plant diseases.
9. apply as claimed in claim 8, described Plant diseases comprise that white muskmelon melon is rotten, capsicum withers, tomato early epidemic, cucumber anthrax, cotton yellow wither, cucumber grey mold, rice bakanae disease, tomato green grass or young crops are withered, one or more in rice banded sclerotial blight, the black shin of tobacco, Root Rot of Wheat, paddy rice rice blast, gibberella saubinetii, rape sclerotium, soybean root rot, tomato gray mould, cucumber foxiness or cucumber downy mildew.
10. an agricultural chemicals, is characterized in that, described agricultural chemicals comprises Sang Puxun streptomycete bacterial strain according to claim 1 and/or Sang Puxun Streptomyces culture according to claim 7.
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Cited By (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087438A (en) * | 2015-08-17 | 2015-11-25 | 河南科技学院 | Streptomyces eurocidicus, fungicide containing same as well as preparation method and application of streptomyces eurocidicus |
| EP3302068A4 (en) * | 2015-06-08 | 2018-12-12 | Indigo AG, Inc. | Streptomyces |
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