CN109122722A - Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth - Google Patents
Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth Download PDFInfo
- Publication number
- CN109122722A CN109122722A CN201810817273.3A CN201810817273A CN109122722A CN 109122722 A CN109122722 A CN 109122722A CN 201810817273 A CN201810817273 A CN 201810817273A CN 109122722 A CN109122722 A CN 109122722A
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- CN
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- Prior art keywords
- garlic
- bacillus subtilis
- growth
- application
- promotor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
-
- 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
-
- 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
- A01N33/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds
- A01N33/16—Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds containing nitrogen-to-oxygen bonds
- A01N33/18—Nitro compounds
- A01N33/20—Nitro compounds containing oxygen or sulfur attached to the carbon skeleton containing the nitro group
- A01N33/22—Nitro compounds containing oxygen or sulfur attached to the carbon skeleton containing the nitro group having at least one oxygen or sulfur atom and at least one nitro group directly attached to the same aromatic ring system
-
- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/10—Aromatic or araliphatic carboxylic acids, or thio analogues thereof; Derivatives thereof
-
- 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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
- A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring
Abstract
Garlic endophyte bacillus subtilis (Bacillus subtilis) is promoting the application in Garlic Growth, promotes Garlic Growth by spraying bacillus subtilis bacterium solution to garlic plant basal part of stem.The present invention also provides bacillus subtilises to prepare the application in Garlic Growth promotor, including as active constituent bacillus subtilis and carrier, the growth promoter be promote garlic plant height and fresh weight promotor.In addition, the Biostimulation agent further includes compounding agent, the compounding agent is at least one of gibberellin, methyl α-naphthyl acetate, compound sodium nitrophenolate.Promote Garlic Growth, non agricultural chemical residuum with bacillus subtilis, non-environmental-pollution does not cause security threat to people and animals；With Garlic Growth promotor prepared by the fermentation liquid of bacillus subtilis, preparation process is simple, and at low cost, effect is good, easy to use.
Description
Technical field
The present invention relates to biological control and biological pesticides more particularly to garlic endophyte bacillus subtilis to promote garlic
Application in growth.
Background technique
Garlic (Allium sativum L.) is Liliaceae allium crop, is to produce with bulb (garlic), garlic stems and shoot
Product, it is full of nutrition, it is one of most popular conventional modulated product and important health food.China is the main production of garlic
State, country of consumption and exported country, Garlic yield account for about the 90% of global yield, occupy first place in the world.With agricultural restructuring by
Deeply, planting garlic area expands step year by year, and problem continuously emerges, and therefore, how to improve Garlic yield and quality, reduces production
Cost realizes garlic No-harmful apple orchard, and the rational application of fertilizer plays an important role, and at this stage, garlic is applied in production according to the survey
There is many problems for fertilizer, for example blindly fertilising phenomenon is serious, precipitation is uneven, organic fertilizer investment is reduced and excessively applied
The problems such as fertile.Developing novel plant growth promoter is the new direction for solving the problems, such as Garlic Fertilization.
Endophyte of plant is the resource microorganism having a extensive future due to that can generate rich and varied metabolite,
Its important ecological effects in nature, causes the extensive concern of people.Endophyte of plant because with plant symbiosis,
There are adjusting and reparation to the microenvironment of plant growth and surrounding plants.Bud pole bacterium (Bacillus spp.) is application
One of more biocontrol bacteria, because it can generate the endospore of heat-resisting, drought-enduring, uvioresistant and organic solvent, so being
Ideal screening of biocontrol agents object.Bacillus subtilis (Bacillus subtilis) has extensive antimicrobial spectrum, to rice banded sclerotial blight
Disease, notoginseng root rot, tobacco black shank etc. have significant resistance.But bacillus subtilis as plant growth promoter in agriculture
There is not application in industry also.Common plant growth promoter such as gibberellin (GA), methyl α-naphthyl acetate (NAA), compound sodium nitrophenolate etc. all pass through
What chemically synthesized method generated, synthesis cost is high, and it is complicated for operation, and not only cultivated simply using endophyte of plant, it is at low cost,
And effect is good.
Summary of the invention
In order to overcome chemical pesticide environmental pollution present in prevention and treatment garlic white rot, pesticide residue and threaten people and animals strong
The problems such as health, the present invention provides bacillus subtilises (Bacillus subtilis) to promote the application in Garlic Growth, leads to
It crosses to garlic plant basal part of stem sprinkling bacillus subtilis bacterium solution and promotes Garlic Growth, this constitutes the first aspect of the invention.
The present invention also provides bacillus subtilises to prepare the application in Garlic Growth promotor, including as activity at
The bacillus subtilis divided and carrier, the growth promoter are the promotor for promoting garlic plant height and fresh weight.In addition, institute
Stating Biostimulation agent further includes compounding agent, the compounding agent is gibberellin (GA), methyl α-naphthyl acetate (NAA), in compound sodium nitrophenolate at least
One kind, this constitutes the second aspect of the invention.
Beneficial effects of the present invention: with bacillus subtilis promote Garlic Growth, non agricultural chemical residuum, non-environmental-pollution, no
Security threat is caused to people and animals；With Garlic Growth promotor prepared by the fermentation liquid of bacillus subtilis, preparation process is simple, at
This is low, and effect is good, easy to use.
Bacillus subtilis of the invention is Bacillus subtilis subsp.Inaquosorum KCTC 13429
(T).Document source: Yi H, et al.2014.Genomic insights into the taxonomic status of
the three subspecies of Bacillus subtilis.Systematic and applied
microbiology.37:95-99.
Bacillus subtilis Bacillus subtilis KCTC13429 (T) can pass through from the garlic phosphorus stem that various regions are planted
Conventional method separation screening and obtain, the applicant can also be contacted and asked for.Applicant states 20 years from the application applying date
Interior, the public can contact applicant and ask for the strain.
Detailed description of the invention
Fig. 1 is the photo that bacterial strain 113 described in the embodiment of the present invention 1 cultivates bacterium colony for 24 hours on LB plate；
Fig. 2 is the photo that bacterial strain 113 described in the embodiment of the present invention 2 produces auxin (IAA) in vitro；Wherein, left figure
The photo for producing IAA for 113, right figure are LB blank control；
Fig. 3 is the photo that bacterial strain 113 promotes Garlic Growth in the greenhouse described in the embodiment of the present invention 3, wherein left
Figure is the control for not connecing 113 bacterium solution of bacterial strain, and right figure is the processing of 113 bacterium solution of inoculating strain.
Specific embodiment:
Embodiment 1: the bacterial strain identification of bacillus subtilis (Bacillus subtilis) 113
The garlic phosphorus stem for selecting health, extracts endophyte, and isolate and purify by conventional method, numbers.Wherein, number is
The morphological feature of 113 pure bacterial strain is as shown in Figure 1, the well-grown on LB solid plate of bacterial strain 113, is cultivated under the conditions of 37 DEG C
After for 24 hours, bacterium colony be it is light yellow, shape be circle, rough surface, diameter about 2-4mm.Using LB liquid medium in 37 DEG C,
After 200rpm shaking flask culture for 24 hours, with 8000rpm, thalline were collected by centrifugation, by multistage microwave amplifier phage gene group DNA, uses bacterium
Universal primer 27F:5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R:5'-AAGGAGGTGATCCAGCCGCA-3' are carried out
The amplification of 16S rDNNA gene PCR.PCR reaction system are as follows: 10 × buffer Mix (including Taq enzyme) 12.5 μ L, 10 μm of ol/L
0.5 10.5 μ L of μ L, ddH2O of 27F 0.5 μ L, 10 μm of ol/L 1492R, 1 μ L of template DNA, pcr amplification product is through 0.8% fine jade
After sepharose electrophoresis detection, send to Shanghai Sheng Gong Co., Ltd and be sequenced, as a result as shown in SEQ ID No.1.In NCBI and
EzBio Cloud compares sequencing result on website, and comparison result is as shown in table 1, the bacterial strain and bacillus subtilis Bacillus
The similitude of subtilis KCTC13429 (T) bacterial strain is 99.93%.
The sequence alignment result of table 1 113
Embodiment 2: bacterial strain 113 generates the measurement for promoting plant growth substance in vitro
Bacterial strain 113 shakes training in 20mL SMS fluid nutrient medium, revolving speed 200rpm, and 37 DEG C of temperature.After 4 days, it is added
The tryptophan of 0.5mg/mL, is placed at room temperature for.Then it draws 1mL cell suspending liquid and 2mL color developing agent is added into new centrifuge tube, so
Room temperature dark stands 20min afterwards, compares not connect the LB liquid medium of 113 bacterial strains, if compared with the control, connecing bacterium and handling out
Existing pink then shows that 113 bacterial strains produce IAA (Fig. 2), indicates that 113 cannot generate IAA if not occurring.SMS culture medium
It is formulated as follows: 1L: sucrose 10.0g, (NH4)2SO41.0g, K2HPO42.0g；MgSO40.5g；NaCl 0.1g；Yeast extraction
Object 0.5g；CaCO30.5g；PH 7.2. color developing agent (ready-to-use) preparation method: taken in draught cupboard 15mL the concentrated sulfuric acid and
The deionized water of 25mL, the concentrated sulfuric acid is slowly added in deionized water, while being stirred rapidly and (being mixed, heat dissipation) with glass bar；To
After sulfuric acid solution is cooled to room temperature, then plus 0.5mol/L 750 μ L of ferric chloride solution, stand-by after mixing, as Salkowski ' s is aobvious
Toner.
Phos test media is prepared, is connect 113 single bacterium drop points and culture medium center, 30 DEG C of inversion trainings with sterilizing toothpick
It supports, can bacterium surrounding is observed after 3 days generate transparent circle, if there is then illustrating that the bacterial strain has solution Phos activity, record molten
Phosphorus loop diameter and colony diameter, the two ratio reaction solution the active power of Phos, if bacterium colony surrounding cannot generate it is transparent
Circle then shows that the bacterium is active (table 2) without solution Phos.Phos test cultures based formulas is as follows: glucose 10.0g, magnesium sulfate
0.25g, calcium phosphate 5.0g, potassium chloride 0.2g, ammonium sulfate 0.1g, glucose 10.0g, magnesium chloride 5.0g, agar powder 20.0g.
The ability of 2 113 bacterial strain of table generation growth-promoting substance
Embodiment 3: the index determining of 113 living body of bacterial strain promotion Garlic Growth
The 113 bacterium solutions sterile washing 3 times that 37 DEG C of liquid LB are shaken to training 16h, is finally diluted to 10 for bacterium solution with sterile water8
A cell/mL, so that endophyte processing is used.In 21 × 20 × 15cm3Flowerpot in sow garlic cultivar " improvement " phosphorus stem, 25
DEG C illumination 16h/ dark 8h is cultivated 20-25 days, into the garlic bud phosphorus bud idiophase (6 leaf phase).Take the bacterium of 113 bacterial strain of 17mL
Then liquid is covered with soil, for 24 hours, then 25 DEG C of illumination 16h/ are dark for 25 DEG C of dark moisturizings along one circle of garlic plant basal part of stem spraying
8h is cultivated 15 days.Blank control does not connect endophyte.3 tests are repeated in 4 basins of each processing in total.The 15th natural gift after connecing bacterium
The whole strain plant height that Tong Ji not compare and handle, aerial part plant height, root long, aerial part fresh weight, root fresh weight and whole
Strain plant fresh weight.As shown in figure 3, compared with the control with the processed garlic plant of bacterium solution of bacterial strain 113, growing way is luxuriant, and
Blade is broad.Measurement result is as shown in table 3, compared with the control, the plant height of the processed garlic plant of bacterial strain 113, plant
Fresh weight and root fresh weight are all higher than control, show the growth-promoting significant effect of bacterial strain 113.
The measurement of 3 113 pairs of garlic growth-promoting effects of table
Due to the symbiosis of bacillus subtilis and garlic, bacillus subtilis is advantageous to the healthy growth of garlic
, therefore, the growth that can promote garlic on garlic is being sprayed from being separated after endophyte is enriched in garlic.Withered grass gemma
Bacillus can pulverize with nonionic surfactant, such as tween, fatty amine, the mixing of glycerine fatty acid esters using water as medium
Thick flowable suspension is made, is sprayed on crop surface, it is adsorbable that different dispersions is formed in microparticle surfaces, change
The distribution and infiltration of kind plant basal part of stem biological pesticide, increase operation rate.Secondly, can also be with inert filler and a certain amount of dispersion
Agent reaches certain particle size in proportion after mixing well.These powdered pesticides can directly impose on plant basal part of stem, quilt
It absorbs, water-wet profit, dispersion can also be added, be spray applied.Powdered pesticide has the characteristics that storage and transportation is convenient, safe, and packaging material is easy
Processing, it is also safer to plant.It again, can also be with the mixed preparings such as gibberellin, methyl α-naphthyl acetate or compound sodium nitrophenolate to promote effect.
Sequence table
<110>Jiangsu Normal University
<120>garlic endophyte bacillus subtilis is promoting the application in Garlic Growth
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1370
<212> DNA
<213>bacillus subtilis (Bacillus subtilis)
<400> 1
cttgctccct gatgttagcg gcggacgggt gagtaacacg tgggtaacct gcctgtaaga 60
ctgggataac tccgggaaac cggggctaat accggatggt tgtttgaacc gcatggttca 120
aacataaaag gtggcttcgg ctaccactta cagatggacc cgcggcgcat tagctagttg 180
gtgaggtaac ggctcaccaa ggcaacgatg cgtagccgac ctgagagggt gatcggccac 240
actgggactg agacacggcc cagactccta cgggaggcag cagtagggaa tcttccgcaa 300
tggacgaaag tctgacggag caacgccgcg tgagtgatga aggttttcgg atcgtaaagc 360
tctgttgtta gggaagaaca agtaccgttc gaatagggcg gtaccttgac ggtacctaac 420
cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg 480
tccggaatta ttgggcgtaa agggctcgca ggcggtttct taagtctgat gtgaaagccc 540
ccggctcaac cggggagggt cattggaaac tggggaactt gagtgcagaa gaggagagtg 600
gaattccacg tgtagcggtg aaatgcgtag agatgtggag gaacaccagt ggcgaaggcg 660
actctctggt ctgtaactga cgctgaggag cgaaagcgtg gggagcgaac aggattagat 720
accctggtag tccacgccgt aaacgatgag tgctaagtgt tagggggttt ccgcccctta 780
gtgctgcagc taacgcatta agcactccgc ctggggagta cggtcgcaag actgaaactc 840
aaaggaattg acgggggccc gcacaagcgg tggagcatgt ggtttaattc gaagcaacgc 900
gaagaacctt accaggtctt gacatcctct gacaatccta gagataggac gtccccttcg 960
ggggcagagt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta 1020
agtcccgcaa cgagcgcaac ccttgatctt agttgccagc attcagttgg gcactctaag 1080
gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaaatcatc atgcccctta 1140
tgacctgggc tacacacgtg ctacaatgga cagaacaaag ggcagcgaaa ccgcgaggtt 1200
aagccaatcc cacaaatctg ttctcagttc ggatcgcagt ctgcaactcg actgcgtgaa 1260
gctggaatcg ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggccttgt 1320
acacaccgcc cgtcacacca cgagagtttg taacacccga agtcggtgag 1370
Claims (8)
1. garlic endophyte bacillus subtilis (Bacillus subtilis) is promoting the application in Garlic Growth.
2. garlic endophyte bacillus subtilis according to claim 1 is promoting the application in Garlic Growth, feature
It is, is to promote Garlic Growth to the bacterium solution of garlic sprinkling bacillus subtilis.
3. garlic endophyte bacillus subtilis according to claim 2 is promoting the application in Garlic Growth, feature
It is, the spray method of the bacterium solution of the bacillus subtilis is that the bacterium solution of the bacillus subtilis is sprayed at garlic and is planted
Strain basal part of stem.
4. garlic endophyte bacillus subtilis according to claim 2 is promoting the application in Garlic Growth, feature
It is, the spraying time of the bacterium solution of the bacillus subtilis is the sprinkling when garlic plant grows to the bud phosphorus bud idiophase.
5. the bacillus subtilis in claim 1 is preparing the application in Garlic Growth promotor.
6. bacillus subtilis according to claim 5 is preparing the application in Garlic Growth promotor, which is characterized in that
The Garlic Growth promotor is the promotor for increasing garlic plant height and garlic plant fresh weight.
7. bacillus subtilis according to claim 5 prevents and treats the application of the biological pesticide of garlic white rot in preparation,
It is characterized in that, the Garlic Growth promotor includes bacillus subtilis and the carrier as active constituent.
8. bacillus subtilis according to claim 5 prevents and treats the application of the biological pesticide of garlic white rot in preparation,
It is characterized in that, the Garlic Growth promotor further includes compounding agent, and the compounding agent is gibberellin, methyl α-naphthyl acetate and/or compound nitrophenol
Sodium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810817273.3A CN109122722A (en) | 2018-07-24 | 2018-07-24 | Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810817273.3A CN109122722A (en) | 2018-07-24 | 2018-07-24 | Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth |
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| Publication Number | Publication Date |
|---|---|
| CN109122722A true CN109122722A (en) | 2019-01-04 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810817273.3A Pending CN109122722A (en) | 2018-07-24 | 2018-07-24 | Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth |
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2018
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| CN101092599A (en) * | 2007-05-30 | 2007-12-26 | 江苏省农业科学院 | Preventing Bacillus subtilis grown inside diseased plant |
| CN102925394A (en) * | 2012-11-16 | 2013-02-13 | 江苏省农业科学院 | Endophytic bacillus subtilis with disease prevention and growth promotion effects |
| CN107960103A (en) * | 2014-12-29 | 2018-04-24 | Fmc有限公司 | For promoting plant growth and treating the microbial composite and application method of plant disease |
| CN105907658A (en) * | 2016-03-25 | 2016-08-31 | 洛阳理工学院 | Peony endogenous Bacillus subtilis and its separation method and use |
| CN107502570A (en) * | 2017-08-18 | 2017-12-22 | 华中农业大学 | One plant of Biocontrol Bacillus subtilis BJ 1 and its application |
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Application publication date: 20190104 |