CN108949583A - One plant of three torch -19 of Trichoderma asperellum strain and its application - Google Patents
One plant of three torch -19 of Trichoderma asperellum strain and its application Download PDFInfo
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- CN108949583A CN108949583A CN201810849198.9A CN201810849198A CN108949583A CN 108949583 A CN108949583 A CN 108949583A CN 201810849198 A CN201810849198 A CN 201810849198A CN 108949583 A CN108949583 A CN 108949583A
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- trichoderma asperellum
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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
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
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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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- 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
- C12N1/14—Fungi; Culture media therefor
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Abstract
The invention discloses one plant of trichoderma asperellum (Trichoderma asperellum) bacterial strain, three torches -19, it has been deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (referred to as are as follows: CGMCC) on June 14th, 2018, deposit number is CGMCC No.15870, depositary institution address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.The bacterial strain can inhibit its growth to shot hole streptomycete, sour scab streptomycete, Streptomycesalbidoflhaving, play effectively preventing and therapeutic effect to potato scab.
Description
Technical field
The invention belongs to technical field of bioengineering more particularly to trichoderma asperellum for preventing and treating in potato scab microbial inoculum
Application.
Background technique
Potato scab, be it is a kind of endanger horse bell administration tuberous plant disease.It shows as stem tuber surface and subcircular occurs extremely
The filbert scab of unsetting suberification scab shape or patch, it is coarse that hand touches texture.Two kinds of disease symptoms are generally divided into, are net respectively
Line shape scab and breach shape scab.Although usual scab is only limitted to cortex, but killed potato wedge quality and yield can still reduce, intolerant to
Storage, and sick potato appearance is indecency, commodity grade greatly declines, and incurs certain economic loss.In the past in China within the border due to hair
Sick rate is not high, and area of causing harm is little, so not attracting people's attention.But in recent years, on Inner Mongol, Hebei, Shandong and other places
The onset area in area is increasing, and caused economic loss is also considerable.Therefore, there is an urgent need to carry out science to this disease
Prevention and treatment.
According to people for a long time to the separation identification research of pathogenic microorganism, it has been determined that cause the cause of disease of potato scab
Bacterium is mainly scab streptomycete etc..And these pathogenic bacteria belong to conditioned pathogen, i.e., hold under the conditions of soil environment meta-alkalescence
It easily grows and causes potato disease, and then seldom occur in Acid soil environment.
Control method mainly include the following types: (1), select disease-free potato seed；(2), more organic fertilizers or green manure can partially press down
System morbidity；(3), crop rotations in 5 years or more are carried out with Curcurbitaceae, pulse family, liliaceous vegetable；(4) the vegetable plot plantation that selection has been retained, knot
The potato phase meets arid and should water in time；(5), chemoprophylaxis: some chemical pesticides can be used to prevent and treat shot hole；(6) biological control
Method: screening has good inhibitory effect and the strong microorganism of colonization ability to Bacterial Spot Pathogen, during producing potato seed and agriculture
It is applied in the planting process of field, achievees the effect that inhibit growth of pathogenic bacteria.
Summary of the invention
In order to effectively prevent potato scab, an object of the present disclosure provides one plant of trichoderma asperellum (Trichoderma
Asperellum) three torch -19 of bacterial strain are deposited in China Committee for Culture Collection of Microorganisms on June 14th, 2018
Common micro-organisms center (referred to as are as follows: CGMCC), deposit number are as follows: CGMCC No.15870, depositary institution address are as follows: court of Beijing
The institute 3 of positive area's North Star West Road 1.
The second object of the present invention is to provide a kind of microbial bacterial agent, and active constituent is three torch -19 of Trichoderma asperellum strain.
The third object of the present invention is to provide a kind of method for preparing microbial bacterial agent,
The fourth object of the present invention is to provide the application of three torch -19 and its microbial inoculum of bacterial strain, to shot hole streptomycete
(Streptomyces scabiei), sour scab streptomycete (Streptomyces acidiscabies), Streptomycesalbidoflhaving
(Streptomyces albidoflavus) can inhibit its growth, play effectively preventing to potato scab and treatment is made
With.
Detailed description of the invention:
The fungistatic effect figure of Fig. 1 bacterial strain -19 pairs of shot hole streptomycetes 4.0076 of three torches.
The fungistatic effect figure of Fig. 2 bacterial strain -19 pairs of shot hole streptomycetes 4.1765 of three torches.
The fungistatic effect figure of -19 pairs of three torch of Fig. 3 bacterial strain sour scab streptomycetes 4.1789.
The fungistatic effect figure of -19 pairs of Streptomycesalbidoflhavings of three torch of Fig. 4 bacterial strain 4.3598.
- 19 pairs of shot hole streptomycetes 4.0076 of three torch of Fig. 5 bacterial strain, sour scab streptomycete 4.1789, Streptomycesalbidoflhaving
The fungistatic effect figure of 4.3598 3 kinds of hybrid bacterial strains.
Three torch -19 of Fig. 6 bacterial strain and shot hole streptomycete 4.0076, sour scab streptomycete 4.1789, Streptomycesalbidoflhaving
Fungistatic effect figure during 4.3598 3 kinds of hybrid bacterial strains co-cultivations.
Three torch -19 of Fig. 7 bacterial strain grows schematic diagram.
Specific embodiment:
1, acquisition, separation, the identification of bacterial strain
Strain acquires soil sample from Banana in Fujian sick soil, and separation is identified and obtained.
Colonial morphology: it grows fastly, is cultivated 5 days under 25 DEG C of dark conditions, colony diameter 60- in malt extract medium
65mm, white, the later period is in celandine green because producing spore, cotton-shaped；The back side is light brown, no water colo(u)r.Conidium largely generates,
The area Chan Bao is evenly distributed.Conidiophore is tall and big, and wall is smooth or slightly coarse, usually generates branch, wide 2.5- in relative position
5.0um；It is few to produce spore bottle stalk, several on falx or its branch, flask shape neck flexion, 5.0-16.7*2.5-3.5um；
The wide ellipse or subsphaeroidal of conidium, wall is smooth or slightly coarse, 2.1-4.5um；Chlamydospore is common.
Method for identifying molecules: the complete sequence and 28S region sequence fragment analysis of ITS1/5.8SrRNA, ITS2 are accredited as spine spore
Trichoderma (Trichoderma asperellum).
2, fungal culture method:
Three torch -19 of Trichoderma asperellum strain activates 3 times in solid potato culture medium.Then take 1 ring in potato solid
Plating medium, 28 DEG C, shaking table culture 5 days.
3, strain purposes and effect:
(1) shot hole Antagonistic Fungi screening material and method:
Test cause of disease is shot hole streptomycete (Streptomyces scabiei) 4.0076 and 4.1765, sour scab strepto-
Bacterium (Streptomyces acidiscabies) 4.1789, Streptomycesalbidoflhaving (Streptomyces albidoflavus)
4.3598 four kinds.
Murphy juice culture medium (g/L): potato leaching liquid 1L, glucose 20g, KH2PO43g, MgSO4.7H2O 1.5g,
Vitamin B1 is micro, PH6.0, and 15g agar is added in solid medium.
Bacteria culturing method: four kinds of pathogens are cultivated 4-5 days, 28 DEG C on potato culture respectively；Use 0.2ml
Sterile water scrapes spore and mycelium, and certain density pathogen suspension is made；It is required that mycelia is raw when 100ul volume spread plate
Planar surface can be paved with after length.
(2) bacteriostatic test method:
Tetra- kinds of pathogens of 100ul are respectively coated on different plates (PDA), connect reesei spores in plate partial points.28℃
Culture.It is observed after 3 days.By to shot hole streptomycete (Streptomyces scabiei) 4.0076 and 4.1765, sour sore
Scab streptomycete (Streptomyces acidiscabies) 4.1789, Streptomycesalbidoflhaving (Streptomyces
Albidoflavus) 4.3598 4 kinds of tests, discovery fungi can effectively inhibit these pathogenic microorganisms to grow during the growth process.
As a result: fungi is to 4 kinds of pathogens all effective (Fig. 1, Fig. 2, Fig. 3, Fig. 4)
Inhibit Mixed Microbes growth test: shot hole streptomycete (Streptomyces scabies) 4.0076,4.1765, acid
Scab streptomycete (Streptomyces acidiscabies) 4.1789, Streptomycesalbidoflhaving (Streptomyces
Albidoflavus) 4.3598 each 100ul are coated on same PDA plate, connect 10ul trichoderma asperellum in plate partial points.28℃
Culture.It is observed after 2 days.(Fig. 5) result: trichoderma asperellum has inhibitory effect to mixing pathogen.
Co-culture bacteriostatic test method: shot hole streptomycete (Streptomyces scabies) 4.0076,4.1765, acid
Scab streptomycete (Streptomyces acidiscabies) 4.1789, Streptomycesalbidoflhaving (Streptomyces
Albidoflavus) 4.3598 each 100ul and 10ul trichoderma asperellum mixing are coated on PDA plate, 28 DEG C of cultures.It is seen after 2 days
It examines.As a result: trichoderma asperellum and four kinds of pathogens during symbiosis, can preferred growth, the growth of other four kinds of bacterium is by obvious
Inhibit (Fig. 6).
4, the preparation method of microbial bacterial agent:
1) prepared by potato fluid nutrient medium (g/L): potato leaching liquid 1L, glucose 20g；Solid medium is added
15g agar；
2) three torch -19 of Trichoderma asperellum strain activates 3 times in solid potato culture medium；
3) take 1 ring in 50ml potato fluid nutrient medium, 28 DEG C, shaking table culture 2 days；
4) after fermentation liquid is centrifuged 1min clock through 12,000r/min, supernatant is abandoned, precipitating is suspended in the sterile water of same volume,
Prepare cell suspension.
Sequence table
<110>Fujian Sanju Biotechnology Co., Ltd.
<120>one plants of three torches -19 of Trichoderma asperellum strain and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 600
<212> DNA
<213>trichoderma asperellum (Trichoderma asperellum)
<400> 1
ccttcggggt ggctcctaaa ggttacctca ccgacttcgg gtgttacaaa ctctcgtggt 60
gtgacgggcg gtgtgtacaa ggcccgggaa cgtattcacc gcggcatgct gatccgcgat 120
tactagcgat tccagcttca cgcagtcgag ttgcagactg cgatccgaac tgagaacaga 180
tttgtgggat tggcttaacc tcgcggtttc gctgcccttt gttctgtcca ttgtagcacg 240
tgtgtagccc aggtcataag gggcatgatg atttgacgtc atccccacct tcctccggtt 300
tgtcaccggc agtcacctta gagtgcccaa ctgaatgctg gcaactaaga tcaagggttg 360
cgctcgttgc gggacttaac ccaacatctc acgacacgag ctgacgacaa ccatgcacca 420
cctgtcactc tgcccccgaa ggggacgtcc tatctctagg attgtcagag gatgtcaaga 480
cctggtaagg ttcttcgcgt tgcttcgaat taaaccacat gctccaccgc ttgtgcgggc 540
ccccgtcaat tcctttgagt ttcagtcttg cgaccgtact ccccaggcgg agtgcttaat 600
Claims (8)
1. one plant of three torch -19 of trichoderma asperellum (Trichoderma asperellum) bacterial strain, deposit number is CGMCC No.
15870。
2. a kind of microbial bacterial agent, which is characterized in that its active constituent is three torch -19 of Trichoderma asperellum strain.
3. a kind of method for preparing microbial bacterial agent, it is characterised in that:
1) three torch -19 of Trichoderma asperellum strain activates 3 times in solid potato culture medium；
2) take 1 ring in 50ml potato fluid nutrient medium, 28 DEG C, shaking table culture 2 days；
3) after fermentation liquid is centrifuged 1min clock through 12,000r/min, supernatant precipitating is collected respectively, and precipitating is suspended in same volume
In sterile water, cell suspension is prepared.
4. a kind of application of microbial bacterial agent in prevention and treatment potato scab drug.
5. a kind of three torch -19 of Trichoderma asperellum strain is applied in prevention and treatment potato scab drug.
6. application according to claim 3 or 4, it is characterised in that: the potato scab is by shot hole strepto-
Bacterium causes.
7. application according to claim 3 or 4, it is characterised in that: the potato scab is by sour scab strepto-
Bacterium causes.
8. application according to claim 3 or 4, it is characterised in that: the potato scab is Streptomycesalbidoflhaving
Cause.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810849198.9A CN108949583B (en) | 2018-07-28 | 2018-07-28 | Trichoderma asperellum strain Sanju-19 and application thereof |
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| CN201810849198.9A CN108949583B (en) | 2018-07-28 | 2018-07-28 | Trichoderma asperellum strain Sanju-19 and application thereof |
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| CN108949583A true CN108949583A (en) | 2018-12-07 |
| CN108949583B CN108949583B (en) | 2021-05-18 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111727963A (en) * | 2019-03-20 | 2020-10-02 | 宁夏大学 | Carrier microbial inoculum compound for preventing and controlling scab and preparation method thereof |
| CN113604364A (en) * | 2021-09-01 | 2021-11-05 | 贵州省茶叶研究所 | Trichoderma asperellum strain and application thereof |
-
2018
- 2018-07-28 CN CN201810849198.9A patent/CN108949583B/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111727963A (en) * | 2019-03-20 | 2020-10-02 | 宁夏大学 | Carrier microbial inoculum compound for preventing and controlling scab and preparation method thereof |
| CN113604364A (en) * | 2021-09-01 | 2021-11-05 | 贵州省茶叶研究所 | Trichoderma asperellum strain and application thereof |
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| Publication number | Publication date |
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| CN108949583B (en) | 2021-05-18 |
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