CN107164285A - One bacillus subtilis BBD012 and preventing and treating tomato disease in application - Google Patents
One bacillus subtilis BBD012 and preventing and treating tomato disease in application Download PDFInfo
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- CN107164285A CN107164285A CN201710562201.4A CN201710562201A CN107164285A CN 107164285 A CN107164285 A CN 107164285A CN 201710562201 A CN201710562201 A CN 201710562201A CN 107164285 A CN107164285 A CN 107164285A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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Abstract
The present invention relates to a bacillus subtilis BBD012, Classification And Nomenclature isBacillus subtilis, deposit number：M 2017097；The bacillus subtilis BBD012 can be used for suppressing botrytis cinerea and/or Fulvia fulva.Said preparation is bacillus subtilis BBD012 nutrient solutions, and preparation method is that inoculation bacillus subtilis BBD012 single bacteriums are fallen within LB fluid nutrient mediums, 160 rmin in 28 DEG C of constant-temperature shaking incubator‑124 h are cultivated, using sterilized water by the diluting colonies of gained bacterium solution after culture to 4 × 108 cfu·mL–1, prevention and controls are that tomato plant is sprayed using bacillus subtilis BBD012 nutrient solutions.Bacillus subtilis BBD012 has good antagonistic effect to botrytis cinerea and Fulvia fulva Ciferri, and the prevention effect to graw mold of tomato and leaf mold respectively reaches 64.9% and 58.0%, with good protection effect.
Description
Technical field
The present invention relates to microbial technology field, specifically a kind of bacillus subtilis BBD012 and preventing and treating tomato disease
Application in evil.
Background technology
Graw mold of tomato is that heavier and common disease is endangered on tomato, and each vegetable-growing area all occurs.In addition to tomato is endangered, also
The various crop of harmful eggplant, capsicum, cucumber, bottle gourd etc. 20 can be endangered.Time harm more than low temperature, continuous rainy weather is serious.Hair
Cause cauline leaf withered and substantial amounts of etching, decayed fruit when sick serious, directly affect yield.Graw mold of tomato turns into China's protecting field
The Major Diseases of tomato production, gray mold is to infect institute by Deuteromycotina fungi Botrytis cinerea (Botrytis cinerea)
Cause.
Leaf muld of tomato is by Fulvia fulva（Fulvia fulva）Caused by infecting, in the leaf of tomato, stem, flower, fruit
On, symptom can all occur, but common sympton is that occur on blade, occurs some in vacuum side of blade initial stage and moves back green statin, later stage
It is changed into the mould layer of the irregular shape of grey or black purple, face of blade moves back green flavescence at corresponding position, when serious, and blade often occurs
It is withered to crispatura.In terms of the order of morbidity, often crept up from plant bottom, temperature from 9 DEG C to 34 DEG C, cause of disease can give birth to
Long development, the optimum temperature of development is 20~25 DEG C.In optimum temperature and humidity larger (relative humidity is more than 80%), only
Need generally to fall ill to first quarter moon for 10 days.
Current tomato breeding for disease-resistance is also without breakthrough, country's preventing and treating graw mold of tomato and relying primarily on of leaf mold
Learn medicament.However, long-term, continuous use makes ash arrhizus bacteria and leaf mold generate the resistance to the action of a drug, cause Agro-chemicals control effect
Decline year by year, and chemical agent residual severe contamination agricultural product and environment, jeopardize human and livestock health.Biological control has become control
The important means of disease processed, and endophytic bacterium has the excellent of uniqueness as the potential resource bacterium of biocontrol of plant disease
Gesture, and have shown that good Utilization prospects.Bacillus subtilis (Bacillus subtilis)It is a kind of mesophilous
Aerobic sporiferous Gram-positive bacillus, can produce the interior raw spore of heat-resisting, drought-enduring, uvioresistant and organic solvent
Son, the scope that it suppresses phytopathogen is very wide, including fruit disease after root disease, trunk disease, leaf, flower portion disease and harvest
Evil, is a kind of preferable Biocontrol microorganism.
The content of the invention
It is an object of the present invention to provide a bacillus subtilis BBD012, Classification And Nomenclature is bacillus subtilis
（Bacillus subtilis）BBD012, deposit number is CCTCC NO：M 2017097.
It is a further object of the present invention to provide one kind preventing and treating graw mold of tomato and/or leaf mold preparation.
The bacillus subtilis BBD012 can be used for suppressing botrytis cinerea and Fulvia fulva.
The preventing and treating graw mold of tomato and/or leaf mold preparation are bacillus subtilis BBD012 nutrient solutions.
Contain bacterium colony 4 × 10 in the bacillus subtilis BBD012 nutrient solutions8 cfu·mL– 1。
The preparation method of the bacillus subtilis BBD012 nutrient solutions is：It is inoculated with bacillus subtilis BBD012 single bacterium colonies
In LB fluid nutrient mediums, 160 rmin in 28 DEG C of constant-temperature shaking incubator-124 h are cultivated, after being cultivated using sterilized water
The diluting colonies of gained bacterium solution are to 4 × 108 cfu·mL– 1。
The method of the preparation for treating and prevention graw mold of tomato and/or leaf mold, using bacillus subtilis BBD012
Nutrient solution is sprayed to tomato plant, i.e., bacillus subtilis BBD012 nutrient solutions are sprayed on to the positive and negative of tomato leaf.
Beneficial effect
The bacillus subtilis BBD012 of the present invention is screened from Semen Lablab Album plant, and the bacterial strain is in the mistake with host plant coevolution
Cheng Zhong, forms stable ecological relationship, and endogenetic bacteria can produce the thing of some antibacterials by itself generation or induction host
Matter, or by playing preventive and therapeutic effect to plant disease with phytopathogen competition ecological niche and nutriment.Bacillus subtilis
BBD012 has good antagonistic effect to botrytis cinerea and Fulvia fulva Ciferri, and the bacterial strain is to three kinds of sweet osmanthus germs（Acanthosphere shell,
Grape seat chamber bacterium, anthrax bacteria）Also there is certain antagonism, confirm bacillus subtilis BBD012 bacterium to tomato by testing
Gray mold and the prevention effect of leaf mold respectively reach 64.9% and 58.0%, with good protection effect.
The preservation of biomaterial
One bacillus subtilis BBD012, Classification And Nomenclature is bacillus subtilis（Bacillus subtilis）BBD012,
China typical culture collection center is preserved in, depositary institution is referred to as CCTCC, and preservation address is Wuhan, China Wuhan University,
Deposit number is CCTCC NO：M 2017097, preservation date is on March 8th, 2017.
Brief description of the drawings
Fig. 1 is fungistatic effect figures of the bacillus subtilis BBD012 of the present invention to Fulvia fulva；
Fig. 2 is figures of the bacillus subtilis BBD012 of the present invention to botrytis cinerea fungistatic effect；
In figure：A left side is control, and the right side is fungistatic effect.
Embodiment
Embodiment 1
Potted tomato preventing and treating is carried out using the preparation of prevention graw mold of tomato：
Step 1: inoculation bacillus subtilis BBD012 single bacteriums are fallen within 200mL LB fluid nutrient mediums, 28 DEG C of constant temperature oscillation
160r.min in incubator-124h is cultivated, the bacterium solution cultivated is diluted to 4 × 108cfu.mL– 1, prepare prevention tomato ash
The preparation of mildew；
Step 2: take long to 5-6 leaves, the consistent potted tomato of growing way, it is sprayed to kind with graw mold of tomato preparation is treated and prevented
Eggplant leave dual sides, using clear water as control, by the spore suspension 1 × 10 of botrytis cinerea after 24 h7Individual spore/mL
（The injection of a little sterilized water has been covered with the PDA plate of ash arrhizus bacteria, the spore of agar surface is scraped, sterile chamber is poured into
It is interior, with the filtered through gauze of sterilizing fully after vibration, counted in blood counting chamber）Spray inoculation is thereon.Tomato plant is incubated at
Growth cabinet（23 DEG C, r≤90%）In.3 basins of each processing, per 3 plants of basin, are repeated 3 times.7d investigates the morbidity of tomato after inoculation
Situation, severity Scaling takes 5 grades of standards.0 grade：Disease-free spot, blade is asymptomatic, plant health；1 grade：Lesion area accounts for total blade
Less than 10%；2 grades：Lesion area accounts for the 10%-50% of total blade；3 grades：Lesion area accounts for more than the 50% of total blade；4 grades：Plant
Strain is dead.
Disease index=100 × [Σ（The sick number of sheets × disease number of levels at different levels）]/（Investigate total number of sheets × 4）Prevention effect（%）
=[（Compare disease index after disease index-antagonistic strain processing）]/control disease index] × 100
As a result show：The tomato plant that spraying treats and prevents graw mold of tomato preparation is reduced than the disease index handled with clear water
23.1, prevention effect is 64.9%（It is shown in Table 1）.
Prevention effects of the bacillus subtilis BBD012 of table 1 to graw mold of tomato
Embodiment 2
Potted tomato preventing and treating is carried out using the preparation of prevention leaf muld of tomato：
Step 1: inoculation bacillus subtilis BBD012 single bacteriums are fallen within 200mL LB fluid nutrient mediums, 28 DEG C of constant temperature oscillation
160r.min in incubator-124h is cultivated, the bacterium solution cultivated is diluted to 4 × 108cfu.mL– 1, prepare prevention tomato ash
The preparation of mildew；
Step 2: take long to 5-6 leaves, the consistent potted tomato of growing way, it is sprayed to kind with graw mold of tomato preparation is treated and prevented
Eggplant leave dual sides, using clear water as control, by the spore suspension 1 × 10 of botrytis cinerea after 24 h7Individual spore/mL
（The injection of a little sterilized water has been covered with the PDA plate of ash arrhizus bacteria, the spore of agar surface is scraped, sterile chamber is poured into
It is interior, with the filtered through gauze of sterilizing fully after vibration, counted in blood counting chamber）Spray inoculation is thereon.Tomato plant is incubated at
Growth cabinet（23 DEG C, r≤90%）In.3 basins of each processing, per 3 plants of basin, are repeated 3 times.7d investigates the morbidity of tomato after inoculation
Situation, severity Scaling takes 5 grades of standards.0 grade：Disease-free spot, blade is asymptomatic, plant health；1 grade：Lesion area accounts for total blade
Less than 10%；2 grades：Lesion area accounts for the 10%-50% of total blade；3 grades：Lesion area accounts for more than the 50% of total blade；4 grades：Plant
Strain is dead.
Disease index=100 × [Σ（The sick number of sheets × disease number of levels at different levels）]/（Investigate total number of sheets × 4）Prevention effect（%）
=[（Compare disease index after disease index-antagonistic strain processing）]/control disease index] × 100
As a result show：The tomato plant of inoculation Fulvia fulva is than the disease index that is handled with clear water after inoculation BBD012 bacterial strains
Reduction by 17.7, prevention effect is 58.0%（It is shown in Table 2）.
Table 2：Prevention effects of the bacillus subtilis BBD012 to leaf muld of tomato
Embodiment 3
Screening, separation and the purifying of bacillus subtilis BBD012 bacterial strains:
Step 1: the preparation of culture medium：
Bacterial Plate culture LB culture mediums（Tryptone 10g/L；Yeast extract 5g/L；Sodium chloride 10g/L；Agar powder
18g/L）, Liquid Culture LB fluid nutrient mediums（It is not added with agar powder）, fungal culture and flat board dual test PDA culture medium
（Potato 200g/L；Glucose 20g/L；Agar powder 18g/L）.
Step 2: the blade 3g of the healthy Semen Lablab Album plant of collection, soaks 30s in 75% alcohol, then with 1% sodium hypochlorite
3min is sterilized, after being rinsed 5 times with sterile distilled water, takes the μ L of flushing liquor 200 of last time to be coated on LB culture medium flat plates；Disappear
Blade after poison is ground in sterile mortar, is added 5ml sterile distilled waters and is mixed, takes the μ L of lapping liquid 200, is coated on LB cultures
On base flat board.Cultivate, observe day by day at 28 DEG C, without bacterium colony in the culture medium that such as last time flushing liquor is coated with, lapping liquid is applied
The bacterium colony grown in the culture medium of cloth is endophyte, carries out after purifying culture, is saved backup in 4 DEG C of refrigerators.
Step 3: antagonistic activity is determined
Botrytis cinerea, Fulvia fulva, sweet osmanthus leaf spot fungi (acanthosphere shell), sweet osmanthus anthrax bacteria and sweet osmanthus leaf spot fungi
（Grape seat chamber bacterium）Activation culture, the bacteria cake that 8mm is made is positioned at PDA culture medium center, surrounding 40mm placement four respectively
8mm sizes contain the filter paper of endogenetic bacteria, are placed in culture 4 ~ 5 days in 28 DEG C of incubators, measure inhibition zone radius and calculate suppression
Bacterium rate.
As a result show and 19 plants of endogenetic bacterias are isolated from the blade of healthy Semen Lablab Album, wherein isolated BBD012
Bacteriostatic activity is most strong, to Fulvia fulva bacteriostasis rate up to 65.32%, Fig. 1；To botrytis cinerea bacteriostasis rate up to 79.52%, scheme
2；Also there are a certain degree of inhibitory action, table 3 to other three kinds of pathogens.
Bacteriostasis rate of the bacterial strain BBD012 zymotic fluids of table 3 to 5 kinds of disease funguses
Step 4: the identification of strong Antagonistic Endophytic strain
1st, form and Physiology and biochemistry identification
Method is referred to《Primary Jie Shi Bacteria Identifications handbook》(Buchanan ＆ Gibbons, 1984) and《Common bacteria system is reflected
Determine handbook》(east show pearl and Cai Miaoying, 1999).
1）Morphological feature
Bacterial strain BBD012 cells are shaft-like, Dan Sheng, flagellum Zhousheng, Gram-positive.Bacterium colony is white, opaque, edge
Irregularly.
2）Physiological and biochemical property
Bacterial strain BBD012 is aerobic bacteria, it is impossible to moved, it is possible to use sucrose, fructose, glucose, maltose, mannitol, but can not
, can be with hydrolysis starch and gelatin, decomposable asymmetric choice net milk production acid, citrate test, V-P experiments, nitrate reduction examination using lactose
Test and be positive with catalase test, Phenylalanine dehydrogenase experiment, methyl red test and oxidase test are negative, the most suitable growth
PH7.0,30 DEG C of optimum growth temperature.
2nd, 16S rDNA and gyrA gene sequencings
With bacterial genomes DNA Rapid extraction kits（Shanghai Sheng Gong bio-engineering corporations REF B518225-0050）Extract
Bacterial strain BBD012 genomic DNAs be template.16S rDNA PCR amplifications are using primer 16SL： 5'-
ACGGCTACCTTGTTACGACCT-3'（Its sequence such as SEQ ID NO:1）With primer 16SR： 5'-
AGAGTTTGATCCTGGCTCAG-3' （Its sequence such as SEQ ID NO:2）.The amplification of gyrA gene PCRs is using primer gyrA-f：
CGATCAGGAAATGCGTACGTCCTT（Its sequence such as SEQ ID NO:3）With primer gyrA-r：
CAAGGTAATGCTCCAGGCATTGCT（（Its sequence such as SEQ ID NO:4））.Reaction system (50 μ L)：10×buffer
5. 0μL、dNTPs ( 2. 5 mmol·L-1) 4. 0 μ L, primer 16SR (10 pmolL-1) 2. 0 μ L, primer 16SL
( 10pmol·L-1) 2. 0 μ L, Taq enzyme (5 U μ L-1) 0. 5 μ L, template DNA (50 ng μ L-1) 2μL、ddH2O
34. 5μL.PCR response procedures:94 DEG C of 5 min, 94 DEG C of 1 min, 50 DEG C of 1 min, 72 DEG C of 2 min, 30 circulations；72
℃ 10 min.By PCR primer it is purified after serve Hai Sheng works bio-engineering corporation sequencing.It is soft with Blast according to sequencing result
Part recalls the 16S rDNA sequences and gyrA gene orders of related strain from GenBank databases, and ClustalW is used respectively
V1. 82 softwares carry out Multiple Sequence Alignment, carry out sequence analysis with the softwares of Mega 4. 1 afterwards, using adjacent method (
Neighbor-joining, NJ method) carry out Phylogenetic Analysis and tetraploid rice.
As a result show：Bacterial strain BBD012 16S rDNA（Gene order such as SEQ ID NO：Shown in 5）With gyrA gene sequences
Row（Such as SEQ ID NO：Shown in 6）WithBacillus subtilisHomology is up to 99%.With reference to its morphological feature, physiology
Biochemical characteristic, 16S rDNA and gyrA gene sequencings, the bacterial strain is accredited asB. subtilis BBD012。
SEQUENCE LISTING
<110>University Of Science and Technology Of He'nan
<120>One bacillus subtilis BBD012 and preventing and treating tomato disease in application
<130> 1
<160> 6
<170> PatentIn version 3.3
<210> 1
<211> 21
<212> DNA
<213>Artificial sequence
<400> 1
acggctacct tgttacgacc t 21
<210> 2
<211> 19
<212> DNA
<213>Artificial sequence
<400> 2
agagtttgat cctggctca 19
<210> 3
<211> 24
<212> DNA
<213>Artificial sequence
<400> 3
cgatcaggaa atgcgtacgt cctt 24
<210> 4
<211> 24
<212> DNA
<213>Artificial sequence
<400> 4
caaggtaatg ctccaggcat tgct 24
<210> 5
<211> 1515
<212> DNA
<213>Bacillus subtilis
<400> 5
tagagtttga tcctggctca ggacgaacgc tggcggcgtg cctaatacat gcaagtcgag 60
cggacagatg ggagcttgct cccggatgtt agcggcggac gggtgagtaa cacgtgggta 120
acctgcctgt aagactggga taactccggg aaaccggggc taataccgga tggttgtttg 180
aaccgcatgg ttcaaacata aaaggtggct tcggctacca cttacagatg gacccgcggc 240
gcattagcta gttggtgagg taacggctca ccaaggcgac gatgcgtagc cgacctgaga 300
gggtgatcgg ccacactggg actgagacac ggcccagact cctacgggag gcagcagtag 360
ggaatcttcc gcaatggacg aaagtctgac ggagcaacgc cgcgtgagtg atgaaggttt 420
tcggatcgta aagctctgtt gttagggaag aacaagtacc gttcgaatag ggcggtacct 480
tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg gtaatacgta 540
ggtggcaagc gttgtccgga attattgggc gtaaagggct cgcaggcggt ttcttaagcc 600
tgatgtgaaa gcccccggct caaccgggga gggtcattgg aaactgggga acttgagtgc 660
agaagaggag agtggaattc cacgtgtagc ggtgaaatgc gtagagatgt ggaggaacac 720
cagtggcgaa ggcgactctc tggtctgtaa ctgacgctga ggagcgaaag cgtggggagc 780
gaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa gtgttagggg 840
gtttccgccc cttagtgctg cagctaacgc attaagcact ccgcctgggg agtacggtcg 900
caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc atgtggttta 960
attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaat cctagagata 1020
ggacgtcccc ttcgggggca gagtgacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt 1080
gagatgttgg gttaagtccc gcaacgagcg caacccttga tcttagttgc cagcattcag 1140
ttgggcactc taaggtgact gccggtgaca aaccggagga aggtggggat gacgtcaaat 1200
catcatgccc cttatgacct gggctacaca cgtgctacaa tggacagaac aaagggcagc 1260
gaaaccgcga ggttaagcca atcccacaaa tctgttctca gttcggatcg cagtctgcaa 1320
ctcgactgcg tgaagctgga atcgctagta atcgcggatc agcatgccgc ggtgaatacg 1380
ttcccgggcc ttgtacacac cgcccgtcac accacgagag tttgtaacac ccgaagtcgg 1440
tgaggtaacc ttttaggagc cagccgccga aggtgggaca gatgattggg gtgaggtcgt 1500
aacaaggtag ccgta 1515
<210> 6
<211> 949
<212> DNA
<213>Bacillus subtilis
<400> 6
catgagcgtt attgtgtctc gtgctcttcc agatgttcgt gacggtttaa aaccggttca 60
cagacggatt ttatacgcaa tgaatgattt gggcatgaca agtgacaaac cttataaaaa 120
atccgcgcgt atcgttggag aagttatcgg gaaataccac ccgcacggtg attcagcggt 180
atatgaatcc atggtcagaa tggctcagga tttcaactac cgttatatgc tcgttgacgg 240
tcacggaaac ttcggttctg ttgacggaga ctcagcggcg gccatgcgtt atacagaagc 300
aagaatgtct aaaatctcaa tggagattct tcgtgacatc acaaaagaca caatcgatta 360
ccaggataac tatgacgggt cagaaagaga acctgtcgtt atgccttcaa ggttcccgaa 420
tcttctcgtg aacggtgctg ccggcattgc ggtaggtatg gcaacaaaca ttcctccgca 480
ccagctggga gaaatcattg acggtgtact tgctgtcagt gagaatccgg acattacaat 540
tccagagctt atggaagtca ttccagggcc tgatttcccg accgcgggtc aaatcttggg 600
acgcagcggt atccggaaag catacgaatc aggccgaggc tctatcacga tccgggcaaa 660
agctgagatc gaacaaacat cttcgggtaa agaaagaatt atcgttacag agttacctta 720
ccaagtaaat aaggcgaaat taattgagaa aattgccgat ctcgtaaggg acaaaaagat 780
agagggtatc acagatctgc gtgatgagtc agatcgtaca ggtatgagaa ttgtcattga 840
aatcagacgc gacgccaatg caaatgtcat cttaaacaat ctgtacaaac aaactgctct 900
acaaacatct tttggcatca acctgcttgc acttgtgatg ccagccgaa 949
Claims (10)
1. a bacillus subtilis BBD012, it is characterised in that：Classification And Nomenclature is bacillus subtilis（Bacillus subtilis）, China typical culture collection center has been preserved in, depositary institution is referred to as CCTCC, and deposit number is CCTCC
NO：M 2017097, preservation date is on March 8th, 2017.
2. applications of the bacillus subtilis BBD012 as claimed in claim 1 in botrytis cinerea is suppressed.
3. applications of the bacillus subtilis BBD012 as claimed in claim 1 in Fulvia fulva is suppressed.
4. applications of the bacillus subtilis BBD012 as claimed in claim 1 in preventing and treating graw mold of tomato and/or leaf mold.
5. bacillus subtilis BBD012 as claimed in claim 4 answering in preventing and treating graw mold of tomato and/or leaf mold preparation
With.
6. graw mold of tomato and/or leaf mold preparation are prevented and treated as claimed in claim 5, it is characterised in that：For bacillus subtilis
BBD012 nutrient solutions.
7. graw mold of tomato and/or leaf mold preparation are prevented and treated as claimed in claim 6, it is characterised in that：The bacillus subtilis
Contain bacterium colony 4 × 10 in bacterium BBD012 nutrient solutions8 cfu·mL– 1。
8. bacillus subtilis BBD012 as claimed in claim 7 is treating and preventing graw mold of tomato preparation, it is characterised in that：
The preparation method of the bacillus subtilis BBD012 nutrient solutions is：Inoculation bacillus subtilis BBD012 single bacteriums fall within LB liquid
In culture medium, 160 rmin in 28 DEG C of constant-temperature shaking incubator-124 h are cultivated, gained bacterium solution after being cultivated using sterilized water
Diluting colonies to 4 × 108 cfu·mL– 1。
9. using the method for one of the right 6-8 preparation for treating and prevention graw mold of tomato and/or leaf mold, its feature
It is：Tomato plant is sprayed using bacillus subtilis BBD012 nutrient solutions.
10. the method for graw mold of tomato and/or leaf mold is treated and prevented as described in right 9, it is characterised in that：By withered grass gemma
Bacillus BBD012 nutrient solutions are sprayed on the positive and negative of tomato leaf.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109156477A (en) * | 2018-09-14 | 2019-01-08 | 河北省农林科学院植物保护研究所 | Application of the bacillus subtilis BAB-1 in prevention and treatment leaf muld of tomato |
| CN110511893A (en) * | 2019-08-29 | 2019-11-29 | 湖南科技大学 | Bacillus biocontrol bacterial strain and its application |
| CN111748496A (en) * | 2020-07-03 | 2020-10-09 | 天津坤禾生物科技集团股份有限公司 | Application of Brevibacillus laterosporus MES818 in tomato cultivation |
| CN113564085A (en) * | 2021-08-18 | 2021-10-29 | 潍坊科技学院 | Bacillus subtilis and application thereof in prevention and treatment of eggplant phomopsis |
| 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 |
| CN115927051A (en) * | 2022-07-21 | 2023-04-07 | 东北农业大学 | Biocontrol bacteria, biocontrol compound microbial inoculum and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101928678A (en) * | 2009-06-19 | 2010-12-29 | 陈秀蓉 | Bacillus subtilis B2 microbial agent, compound microbial agent and application |
| CN103103149A (en) * | 2013-01-11 | 2013-05-15 | 陈秀蓉 | Bacillus subtilis S001, application of bacillus subtilis S001, microbial preparation and preparation method of microbial preparation |
| KR20130134875A (en) * | 2012-05-31 | 2013-12-10 | (주)현농 | Culture medium for preventing disease and promoting growth of plant, manufacturing method of microbial agent using the same |
-
2017
- 2017-07-11 CN CN201710562201.4A patent/CN107164285B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101928678A (en) * | 2009-06-19 | 2010-12-29 | 陈秀蓉 | Bacillus subtilis B2 microbial agent, compound microbial agent and application |
| KR20130134875A (en) * | 2012-05-31 | 2013-12-10 | (주)현농 | Culture medium for preventing disease and promoting growth of plant, manufacturing method of microbial agent using the same |
| CN103103149A (en) * | 2013-01-11 | 2013-05-15 | 陈秀蓉 | Bacillus subtilis S001, application of bacillus subtilis S001, microbial preparation and preparation method of microbial preparation |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109156477A (en) * | 2018-09-14 | 2019-01-08 | 河北省农林科学院植物保护研究所 | Application of the bacillus subtilis BAB-1 in prevention and treatment leaf muld of tomato |
| CN110511893A (en) * | 2019-08-29 | 2019-11-29 | 湖南科技大学 | Bacillus biocontrol bacterial strain and its application |
| CN110511893B (en) * | 2019-08-29 | 2022-05-17 | 湖南科技大学 | Bacillus biocontrol strain and application thereof |
| CN111748496B (en) * | 2020-07-03 | 2022-05-13 | 天津坤禾生物科技集团股份有限公司 | Application of Brevibacillus laterosporus MES818 in tomato cultivation |
| CN111748496A (en) * | 2020-07-03 | 2020-10-09 | 天津坤禾生物科技集团股份有限公司 | Application of Brevibacillus laterosporus MES818 in tomato cultivation |
| CN113564085A (en) * | 2021-08-18 | 2021-10-29 | 潍坊科技学院 | Bacillus subtilis and application thereof in prevention and treatment of eggplant phomopsis |
| CN113564085B (en) * | 2021-08-18 | 2021-12-21 | 潍坊科技学院 | Bacillus subtilis and application thereof in prevention and treatment of eggplant phomopsis |
| CN115011505A (en) * | 2022-04-24 | 2022-09-06 | 山东省农业科学院 | Bacillus subtilis and application thereof |
| CN115011505B (en) * | 2022-04-24 | 2023-04-18 | 山东省农业科学院 | Bacillus subtilis and application thereof |
| CN115927051A (en) * | 2022-07-21 | 2023-04-07 | 东北农业大学 | Biocontrol bacteria, biocontrol compound microbial inoculum and application thereof |
| CN115927051B (en) * | 2022-07-21 | 2024-05-07 | 东北农业大学 | Biocontrol bacterium, biocontrol compound microbial inoculum 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 |
| 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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