CN110195033A - One bacillus subtilis and the application in the prevention and treatment of S. lycopersici leaf spot - Google Patents
One bacillus subtilis and the application in the prevention and treatment of S. lycopersici leaf spot Download PDFInfo
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- CN110195033A CN110195033A CN201910486419.5A CN201910486419A CN110195033A CN 110195033 A CN110195033 A CN 110195033A CN 201910486419 A CN201910486419 A CN 201910486419A CN 110195033 A CN110195033 A CN 110195033A
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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
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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
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
Abstract
The invention discloses a bacillus subtilis and the applications in the prevention and treatment of S. lycopersici leaf spot.The present invention provides bacillus subtilis (Bacillus subtilis) ZF161, are CGMCC No.16776 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.Bacillus subtilis ZF161 provided by the invention is one plant of tomato rhizospheric microflora of soil, environmentally friendly to people, animal, Crop securify.The bacterial strain has strong antagonism to S. lycopersici leaf spot.Biological control is carried out to S. lycopersici leaf spot using the bacterial strain, it is a kind of S. lycopersici leaf spot biological control mode of efficient stable, it is easy to act on tomato plant it makes use of the bacterial strain and has the characteristics that strong antagonism to S. lycopersici leaf spot fungi, can preferably meets the requirement for preventing and treating S. lycopersici leaf spot in agricultural production.
Description
Technical field
The present invention relates to technical field of plant disease biological control, in particular to a bacillus subtilis and crawl in tomato
Application in the mould leaf spot prevention and treatment of handle.
Background technique
S. lycopersici leaf spot can be crawled handle mould (Stemphylium solani Weber) and S. lycopersici by eggplant
(Stemphylium lycopersici (Enjoji) Yamamoto) causes, and mainly being crawled by eggplant, handle is mould to infect caused one kind
Serious disease causes tomato leaf, vines and fruit large area necrosis, leads to the tomato underproduction.Currently, being directed to S. lycopersici
Leaf spot mainly uses chemical prevention and control method.However, since the drug resistance of S. lycopersici leaf spot and chemical prevention are to ring
The problems such as border is seriously polluted, the biological control research for carrying out S. lycopersici leaf spot are of great significance.
Currently, although chemical bactericide is still occupied an leading position in terms of control of plant disease, bacillus category biological and ecological methods to prevent plant disease, pests, and erosion
Microbial inoculum is the novel biocontrol bacterium of most development potentiality because its advantage such as efficient, with strong points, nontoxic and pollution-free is gradually taken seriously
Strain.
Summary of the invention
For the deficiency for improving existing Prevention Technique, the purpose of the present invention is to provide a kind of bacillus subtilis and its answer
With the bacillus subtilis has significant antagonism to S. lycopersici leaf spot.
In a first aspect, a claimed bacillus subtilis.
Present invention bacillus subtilis claimed is specially bacillus subtilis (Bacillus subtilis)
ZF161 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation on November 26th, 2018
Number be CGMCC No.16776.
Second aspect, a kind of claimed microorganism formulation.
Present invention microorganism formulation claimed contains the bacillus subtilis (Bacillus subtilis)
ZF161。
Further, the microorganism formulation can be biological prevention and control agent.
Further, the biological prevention and control agent can by the bacillus subtilis (Bacillus subtilis) ZF161 and
Auxiliary agent is formulated.Bacillus subtilis (Bacillus subtilis) ZF161 in the biological prevention and control agent effectively containing
Amount can be 1.0 × 107~1.0 × 109Cfu/mL, such as 1.0 × 108cfu/mL。
The third aspect, a kind of claimed bacteria suspension.
Present invention bacteria suspension claimed is the bacillus subtilis (Bacillus subtilis) ZF161's
Bacteria suspension, wherein the content of the bacillus subtilis (Bacillus subtilis) ZF161 is 1.0 × 107~1.0 ×
109cfu/mL。
Further, the content of bacillus subtilis (Bacillus subtilis) ZF161 is 1.0 × 108cfu/
mL。
The third aspect, claimed bacillus subtilis (Bacillus subtilis) ZF161 or described
Microorganism formulation or the bacteria suspension it is following it is any in application:
(A1) S. lycopersici leaf spot is prevented and treated；
(A2) product for preventing and treating S. lycopersici leaf spot is prepared.
Fourth aspect, claimed bacillus subtilis (Bacillus subtilis) ZF161 or described
Microorganism formulation or the bacteria suspension it is following it is any in application:
(B1) inhibit S. lycopersici Leaf blotch pathogeny；
(B2) product for inhibiting S. lycopersici Leaf blotch pathogeny is prepared.
5th aspect, claimed bacillus subtilis (Bacillus subtilis) ZF161 or described
Microorganism formulation or the bacteria suspension it is following it is any in application:
(C1) prevention and treatment is by S. lycopersici leaf spot fungi associated diseases；
(C2) preparation is for preventing and treating by the product of S. lycopersici leaf spot fungi associated diseases.
In above-mentioned 4th and the 5th aspect, the S. lycopersici leaf spot fungi is that eggplant is crawled the mould (Stemphylium of handle
solani Weber)。
6th aspect, a kind of claimed method for preventing and treating S. lycopersici leaf spot.
The method of prevention and treatment S. lycopersici leaf spot provided by the present invention, it may include following steps: by the withered grass bud
Spore bacillus (Bacillus subtilis) ZF161 or the microorganism formulation or the bacteria suspension impose on and (such as spray) tomato plant
It in strain, is colonized the bacillus subtilis (Bacillus subtilis) ZF161 on tomato leaf, and then inhibits tomato
The mould leaf spot of handle of crawling.
Gemma be metabolized opposing stationary, high-output stress-resistance, can long-term preservation, be conducive to exploitation be microbial inoculum.The withered grass gemma
The gemma of bacillus (Bacillus subtilis) ZF161 can obtain with the following method: LB culture medium be utilized, at 28 DEG C
At a temperature of cultivate the bacillus subtilis (Bacillus subtilis) ZF161 2 days, it is a large amount of to generate bacillus subtilises
Gemma.Storage temperature is 22~28 DEG C, such as 21~27 DEG C.
It is prepared by the 7th aspect, claimed bacillus subtilis (Bacillus subtilis) ZF161
Application in the microorganism formulation or the bacteria suspension.
The beneficial effects of the present invention are:
(1) bacillus subtilis (Bacillus subtilis) ZF161 provided by the invention is one plant of tomato rhizosphere soil
Microorganism, it is environmentally friendly to people, animal, Crop securify.
(2) bacillus subtilis (Bacillus subtilis) ZF161 provided by the invention is to S. lycopersici leaf spot
With strong antagonism.The present invention is using bacillus subtilis (Bacillus subtilis) ZF161 to S. lycopersici tikka
Disease carries out biological control, is a kind of S. lycopersici leaf spot biological control mode of efficient stable, it makes use of the withered grass
Bacillus (Bacillus subtilis) ZF161 is easy to act on and have S. lycopersici leaf spot fungi on tomato plant
There is the characteristics of strong antagonism, can preferably meet the requirement for preventing and treating S. lycopersici leaf spot in agricultural production.
Preservation explanation
Strain name: bacillus subtilis
Latin name: Bacillus subtilis
Join the biomaterial (strain) of Ju: ZF161
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on November 26th, 2018
Collection is registered on the books number: CGMCC No.16776
Detailed description of the invention
Fig. 1 is growth colonial morphology of the bacillus subtilis ZF161 on LB solid medium.
Fig. 2 is the 16S rDNA segment of bacillus subtilis ZF161 and the agarose gel electrophoresis figure of gyrB genetic fragment.
Swimming lane 1 is 1417bp (16S rDNA segment)；Swimming lane 2 is 1417bp (16S rDNA segment)；Swimming lane 3 is 991bp (gyrB gene
Segment)；Swimming lane 4 is 991bp (gyrB genetic fragment)；Swimming lane M is molecular weight standard (5000bp).
Fig. 3 is the 16S rDNA phylogenetic tree of bacillus subtilis ZF161.
Fig. 4 is the gyrB Phylogenetic Tree of bacillus subtilis ZF161.
Fig. 5 is S. lycopersici leaf spot fungi and bacillus subtilis ZF161 plate opposite culture result.From left to right according to
It is secondary that (50% iprodione wettable powder is compareed to the inhibitory effect of pathogen, blank control and medicament for bacillus subtilis ZF161
Agent).
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The various culture mediums that following embodiments are related to all are standard mediums commonly used in the art, such as:
LB liquid medium: tryptone 1% (w/v), yeast extract 0.5% (w/v), sodium chloride 1% (w/v),
pH7.0。
LB solid medium (abbreviation LB culture medium) is to be added to agar on the basis of above-mentioned LB liquid medium
1.5%-2% (w/v), as tryptone 1% (w/v), yeast extract 0.5% (w/v), sodium chloride 1% (w/v), agar
1.8% (w/v), pH7.0.
Embodiment 1, the separation screening of bacillus subtilis (Bacillus subtilis) ZF161 and identification
From the rhizosphere soil isolated strains of the Daxing District greenhouse acquisition of continuous plantation tomato for many years.Picking single bacterium is fallen within
It is cultivated in LB liquid medium, obtains bacteria suspension；It is filtered out from the bacterium colony that step (1) is chosen with tablet face-off method and is crawled to tomato
Bacterial strain (table 1) of the mould leaf spot fungi of handle (eggplant crawl handle mould (Stemphylium solani Weber)) with bacteriostasis.Finally
Obtain the bacterial strain that one plant of bacterium number is ZF161.
Table 1 screens Antagonistic bacteria strains to the inhibiting effect of eggplant stemphylium botryosum
| Processing | Concentration | Colony diameter (mm) | Bacteriostasis rate (%) |
| IVF19 | 108cfu/mL | 23.00±0.41 | 70.51±0.93ab |
| IVF43 | 108cfu/mL | 31.67±0.47 | 59.40±1.60d |
| A-5 | 108cfu/mL | 31.00±2.27 | 60.26±5.67d |
| ZF161 | 108cfu/mL | 19.00±1.08 | 75.64±2.60a |
| 18051709-2-1 | 108cfu/mL | 28.67±1.70 | 43.16±4.27cd |
| 18051709-2-2 | 108cfu/mL | 24.67±0.47 | 68.38±1.25bc |
| 18051709-2-3 | 108cfu/mL | 27.67±0.94 | 64.53±2.73bcd |
| 50% iprodione wettable powder | 3g/L | 30.33±0.47 | 61.11±1.57d |
| Blank cultures | 78.00±0.41 |
Note: different lowercases indicate that above there were significant differences in 0.05 level.
Bacterial strain ZF161 morphological feature: ZF161 is gram-positive bacteria, is in elongated rod shape.
On LB plate, for 24 hours, formation bacterium colony is rounded, and milky, edge is irregular, impermeable for 28 DEG C of cultures by bacterial strain ZF161
Bright, there is fold on surface.Fig. 1 is the colonial morphology of bacterial strain ZF161.
The physiological and biochemical property of bacterial strain ZF161: hydrolysis starch, oxidase positive.
The amplification method of the 16S rDNA of bacterial strain ZF161 are as follows: picking single colonie is placed in and shakes bacterium equipped with LB liquid medium
DNA extraction is carried out after cultivating in pipe, carries out PCR amplification by template of DNA.
PCR amplification system is 1 μ L, Mix 10 μ L, upstream and downstream primer each 1 μ L, ddH of 20 μ L:DNA template2O supplies residual body
Product.Amplification condition: 94 DEG C of 5min；94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 90s, 35 circulations；72℃10min.
PCR product is detected through 1% agarose gel electrophoresis, and gel imaging is shown in Figure 2.Gained is expanded through serial analysis
16S rDNA sequence length be 1417bp (SEQ ID No.1).The 16S rDNA sequence is sequenced and carries out BLAST
Analysis is compared, the similarity highest of the 16S rDNA and Bacillus subtilis, have reached 100% as the result is shown.
The amplification method of the gyrB gene of bacterial strain ZF161 are as follows: picking single colonie is placed in and shakes bacterium equipped with LB liquid medium
DNA extraction is carried out after cultivating in pipe, carries out PCR amplification by template of DNA.
PCR amplification system is 1 μ L, Mix 10 μ L, upstream and downstream primer each 1 μ L, ddH of 20 μ L:DNA template2O supplies residual body
Product.Amplification condition: 94 DEG C of 5min；94 DEG C of 30s, 62 DEG C of 30s, 72 DEG C of 1min, 35 circulations；72℃10min.
PCR product is detected through 1% agarose gel electrophoresis, and gel imaging is shown in Figure 2.Gained is expanded through serial analysis
GyrB gene order length be 991bp (SEQ ID No.2).The gyrB gene order is sequenced and carries out BLAST
Analysis is compared, the similarity highest of the gyrB gene and Bacillus subtilis, has reached 98% as the result is shown.
By the 16S rDNA sequence after ncbi database is arranged and compared, systematic evolution tree is constructed.It is shown in Figure 3,
Bacterial strain (label is in Fig. 3) of the invention is most short with the evolutionary distance of B.subtilis, is the approximation of B.subtilis
Kind.
By the gyrB gene order after ncbi database is arranged and compared, systematic evolution tree is constructed.It is shown in Figure 4,
Bacterial strain (label is in Fig. 4) of the invention is most short with the evolutionary distance of B.subtilis, is the approximation of B.subtilis
Kind.
Combining form feature and molecular biology identification finally determine that bacterial strain ZF161 of the invention is bacillus subtilis
(Bacillus subtilis)。
Bacterial strain ZF161 has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC).Ground
Location: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101.Preservation date
On November 26th, 2018.Deposit number is CGMCC No.16776.Strain name: bacillus subtilis；Latin name: Bacillus
subtilis；Join the biomaterial (strain) of Ju: ZF161.
The preparation of embodiment 2, bacillus subtilis (Bacillus subtilis) ZF161 bacteria suspension
By bacillus subtilis (Bacillus subtilis) ZF161 after purification, single colonie is seeded in LB Liquid Culture
It in base, is cultivated 2 days under the conditions of 180r/min at 28 DEG C, a large amount of gemma for generating bacillus subtilis.Gained bacteria suspension contains institute
The effective content for stating bacillus subtilis is 107~109cfu/mL。
Embodiment 3, suppression of bacillus subtilis (Bacillus subtilis) ZF161 to S. lycopersici leaf spot fungi
The simple identification of production
By S. lycopersici leaf spot fungi, (eggplant handle of crawling is mould (Stemphylium solani Weber), by Chinese agriculture section
The comprehensive anti-group of offer of vegetable or flower research institute, institute dish disease) it is connected to after PDA plate center cultivates for 24 hours, by the withered of the embodiment of the present invention 1
Careless bacillus (Bacillus subtilis) ZF161 bacteria suspension symmetrically accesses PDA plate, distance center in four horn shapes
2.5cm, is arranged 3 repetitions and blank control, at 28 DEG C, after cultivating 7d, measures colony diameter using crossing method, calculates suppression
Rate processed.
Inhibiting rate=[(control colony diameter-processing colony diameter)/(control colony diameter-stem diameter)] × 100%
It the results are shown in Table 2 and Fig. 5, under blank cultures, S. lycopersici leaf spot fungi fast growing, the bacterium of growth 7 days
Falling diameter is 78mm, and is added in the PDA culture medium of ZF161 bacteria suspension, S. lycopersici leaf spot fungi slow growth, and bacterium colony is straight
Diameter is 19mm, therefore calculating bacteriostasis rate is 75.6%.
Bacteriostasis rate of 2 bacillus subtilis of table (Bacillus subtilis) ZF161 to S. lycopersici leaf spot fungi
Note: 50% iprodione wettable powder is comparison medicament.Different lowercase expressions have significant in 0.05 level
Difference.
Embodiment 4, tomato tooth in vitro on-chip testing bacillus subtilis (Bacillus subtilis) ZF161 kind
Eggplant is crawled the application effect in the mould leaf spot biological control of handle
Test is that excised leaf is tested, and takes 45 leaf tomato leafs, and processing, clear water control are respectively that be placed in bottom aqueous for 15 leaves
Moisture preservation box in.By the bacillus subtilis cultivated in embodiment 2 (Bacillus subtilis), ZF161 bacteria suspension is (about
108Cfu/mL (it is water mist that the standard that sprays, which is on plant, not will form drop and flows down)) is sprayed on tomato leaf by spray-on process,
By S. lycopersici leaf spot fungi, (eggplant is crawled mould (the Stemphylium solani of handle after blade surface dries, then through spray-on process
Weber)) bacteria suspension (concentration 1 × 108Cfu/ml it) is connected on tomato leaf.Moisture preservation box is sealed up after connecing bacterium, is placed in 28 DEG C of training
It supports in case.After the onset of blade is abundant, disease index is counted to clear water control group (substituting ZF161 bacteria suspension with clear water), is calculated anti-
Effect.
Disease survey is using S. lycopersici leaf spot grade scale: 0 grade: disease-free spot；1 grade: lesion area accounts for entire leaf
5% or less area；3 poles: lesion area accounts for the 6%~25% of entire leaf area；5 grades: lesion area accounts for entire leaf area
26%~50%；7 grades: lesion area accounts for the 51%~75% of entire leaf area；9 grades: lesion area accounts for entire leaf area
76% or more.
Disease index=∑ (the values at different levels × sick number of sheets at different levels)/(investigation total number of sheets × superlative degree value) × 100.
Preventive effect (%)=(control group disease index-processing group disease index)/control group disease index × 100.
The results are shown in Table 3.Obtained bacillus subtilis (Bacillus is cultivated using the embodiment of the present invention 2
Subtilis) ZF161 bacteria suspension (about 108Cfu/mL) the S. lycopersici leaf spot fungi handled, disease incidence disease index are
30.86, and it is 94.07 that clear water, which compares lower disease index, embodiment 2 cultivates obtained bacillus subtilis (Bacillus
Subtilis) ZF161 bacteria suspension (about 108Cfu/mL average preventive effect) is 69.83%
3 bacillus subtilis of table (Bacillus subtilis) ZF161 is (in vitro to the preventive effect of S. lycopersici leaf spot
Blade)
Note: 50% iprodione wettable powder is comparison medicament.
Embodiment 5, test bacillus subtilis (Bacillus subtilis) ZF161 crawls in tomato on tomato plant
Application effect in the mould leaf spot biological control of handle
The test is greenhouse pot culture test, as far as possible exclusion external interference condition.Greenhouse is located at Chinese Academy of Agricultural Sciences vegetables
Flowers research institute.Tomato variety is Qi Dali, susceptible to S. lycopersici leaf spot.Processing and control are respectively 30 plants of tomatoes.
When tomato is long to 5 groups of auxiliary leaves, by the bacillus subtilis cultivated in embodiment 2 (Bacillus subtilis) ZF161 bacterium
Suspension (about 108Cfu/mL) be sprayed on tomato plant by spray-on process, for 24 hours after, then by spray-on process by S. lycopersici tikka
Germ (eggplant crawl handle mould (Stemphylium solani Weber)) bacteria suspension (concentration 1 × 108Cfu/ml) it is inoculated into tomato plant
In strain.Control is clear water.It connects and tomato seedling is put into relative humidity 95%, moisturizing culture in 26~28 DEG C of moisturizing cabinets of temperature after bacterium
48h is transferred to normal seedling cultivation greenhouse culture later.It sufficiently falls ill to clear water control group (substituting ZF161 bacteria suspension with clear water) plant
Afterwards, disease index is counted, preventive effect is calculated.
Disease survey is using S. lycopersici leaf spot grade scale with embodiment 4.
Disease index=∑ (the values at different levels × sick number of sheets at different levels)/(investigation total number of sheets × superlative degree value) × 100.
Preventive effect (%)=(control group disease index-processing group disease index)/control group disease index × 100.
The results are shown in Table 4.Utilize the bacillus subtilis (Bacillus cultivated in the embodiment of the present invention 2
Subtilis) ZF161 bacteria suspension (about 108Cfu/mL) the S. lycopersici leaf spot disease incidence disease index handled is 16.93,
And it is 45.44 that clear water, which compares lower disease index, the bacillus subtilis (Bacillus cultivated in embodiment 2
Subtilis) ZF161 bacteria suspension (about 108Cfu/mL average preventive effect) is 63.27%
Preventive effect (plant) of 4 bacillus subtilis of table (Bacillus subtilis) ZF161 to S. lycopersici leaf spot
Note: 50% iprodione wettable powder is comparison medicament.Different lowercase expressions have significant in 0.05 level
Difference.
Present invention is not limited to the embodiments described above, without departing from the essence of the present invention, this field skill
Any deformation, improvement, the replacement that art personnel are contemplated that each fall within the scope of the present invention.
Sequence table
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<120>one bacillus subtilis and the application in the prevention and treatment of S. lycopersici leaf spot
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gtacgtgtta atgttgtttg taaacgagta aatgttgctt gtgtagctgt cattgtattg 660
caaagccact tcaaccgtaa tgccgtcctt ttcgccttca atgtaaatcg gctcttcatg 720
gacaacctct ttagagcggt ttaaatactc tacataactg ttaattccgc cttcgtaatg 780
gtattcattt ttgcgctctt gtccttcacg tttatcttca atcgtgatgt ttacgccctt 840
tgttaaaaag gccaattcac gcacgcggtt ggcaagcaaa atcgaaatca tactcggttg 900
tttctgaaaa aaatttcggg gtccgggaac aaaatgtgcc gccgttcctg gatgatccgt 960
ttcgccaatg atttcaaggg tttgtaaacc g 991
Claims (10)
1. bacillus subtilis (Bacillus subtilis) ZF161, general in China Committee for Culture Collection of Microorganisms
The deposit number at logical microorganism center is CGMCC No.16776.
2. microorganism formulation, it is characterised in that: the microorganism formulation contains bacillus subtilis described in claim 1
(Bacillus subtilis)ZF161。
3. bacteria suspension, it is characterised in that: the bacteria suspension is bacillus subtilis (Bacillus described in claim 1
Subtilis) the bacteria suspension of ZF161, wherein the content of the bacillus subtilis (Bacillus subtilis) ZF161 is
1.0×107~1.0 × 109cfu/mL；
Further, the content of bacillus subtilis (Bacillus subtilis) ZF161 is 1.0 × 108cfu/mL。
4. bacillus subtilis (Bacillus subtilis) ZF161 or as claimed in claim 2 described in claim 1 is micro-
Biological agent or bacteria suspension as claimed in claim 3 it is following it is any in application:
(A1) S. lycopersici leaf spot is prevented and treated；
(A2) product for preventing and treating S. lycopersici leaf spot is prepared.
5. bacillus subtilis (Bacillus subtilis) ZF161 or as claimed in claim 2 described in claim 1 is micro-
Biological agent or bacteria suspension as claimed in claim 3 it is following it is any in application:
(B1) inhibit S. lycopersici leaf spot fungi；
(B2) product for inhibiting S. lycopersici leaf spot fungi is prepared.
6. bacillus subtilis (Bacillus subtilis) ZF161 or as claimed in claim 2 described in claim 1 is micro-
Biological agent or bacteria suspension as claimed in claim 3 it is following it is any in application:
(C1) prevention and treatment is by S. lycopersici leaf spot fungi associated diseases；
(C2) preparation is for preventing and treating by the product of S. lycopersici leaf spot fungi associated diseases.
7. application according to claim 5 or 6, it is characterised in that: the S. lycopersici leaf spot fungi is that eggplant handle of crawling is mould
(Stemphylium solaniWeber)。
8. a kind of method for preventing and treating S. lycopersici leaf spot, includes the following steps: bacillus subtilis described in claim 1
Bacteria suspension described in microorganism formulation described in (Bacillus subtilis) ZF161 or claim 2 or claim 3 imposes on kind
On eggplant plant.
9. according to the method described in claim 8, it is characterized by: being by the bacillus subtilis in the method
(Bacillus subtilis) ZF161 or the microorganism formulation or the bacteria suspension are sprayed on tomato plant.
10. the bacillus subtilis (Bacillus subtilis) ZF161 described in claim 1 is in preparation claim 2
Application in the microorganism formulation or bacteria suspension as claimed in claim 3.
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| CN115011505A (en) * | 2022-04-24 | 2022-09-06 | 山东省农业科学院 | Bacillus subtilis and application thereof |
| CN115369062A (en) * | 2022-09-02 | 2022-11-22 | 上海市农业科学院 | Tomato bacterial wilt antagonistic bacterium WJB0802 and application thereof |
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Cited By (4)
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| CN115011505A (en) * | 2022-04-24 | 2022-09-06 | 山东省农业科学院 | Bacillus subtilis and application thereof |
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| CN115369062B (en) * | 2022-09-02 | 2023-10-20 | 上海市农业科学院 | Tomato bacterial wilt antagonistic bacterium WJB0802 and application thereof |
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| CN110195033B (en) | 2021-03-02 |
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