CN109566656A - Bei Laisi bacillus is promoting the application in Garlic Growth - Google Patents
Bei Laisi bacillus is promoting the application in Garlic Growth Download PDFInfo
- Publication number
- CN109566656A CN109566656A CN201811559654.2A CN201811559654A CN109566656A CN 109566656 A CN109566656 A CN 109566656A CN 201811559654 A CN201811559654 A CN 201811559654A CN 109566656 A CN109566656 A CN 109566656A
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- CN
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- Prior art keywords
- garlic
- bei laisi
- laisi bacillus
- growth
- bacillus
- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- 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
-
- 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
- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
Abstract
The bacterium solution of the Bei Laisi bacillus is sprayed at garlic plant basal part of stem when garlic bulb grows to 6 leaf to promote Garlic Growth by application of the Bei Laisi bacillus (Bacillus velezensis) in promotion Garlic Growth.The present invention also provides Bei Laisi bacillus to prepare the application in Garlic Growth promotor, including as the Bei Laisi bacillus of active constituent and carrier, the growth promoter is the promotor for promoting garlic plant height.Promote Garlic Growth, non agricultural chemical residuum with Bei Laisi bacillus, non-environmental-pollution does not cause damages to people and animals and soil, and with Garlic Growth promotor prepared by the fermentation liquid of Bei Laisi bacillus, 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 Bei Laisi bacillus in promoting Garlic Growth
Using.
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, planting garlic area expand year by year, and developing novel plant growth promoter is the new direction for solving the problems, such as Garlic Fertilization.It plants
Object endophyte is the resource microorganism having a extensive future, in nature due to that can generate rich and varied metabolite
In important ecological effects, cause the extensive concern of people.Endophyte of plant because with plant symbiosis, to plant growth
And the microenvironment of surrounding plants plays the role of adjusting and reparation.Bacillus (Bacillus spp.) is using more biological and ecological methods to prevent plant disease, pests, and erosion
One of bacterium, because it can generate the endospore of heat-resisting, drought-enduring, uvioresistant and organic solvent, so being ideal biological and ecological methods to prevent plant disease, pests, and erosion
Bacterium screens object.Common plant growth promoter such as gibberellin (GA), methyl α-naphthyl acetate (NAA), compound sodium nitrophenolate etc. are all to pass through chemistry
What synthetic 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
Of the existing technology in order to overcome the problems, such as, the present invention provides Bei Laisi bacillus (Bacillus
Subtilis) promoting the application in Garlic Growth, when garlic bulb grows to 6 leaf, by the Bei Laisi bacillus
Bacterium solution is sprayed at garlic plant basal part of stem to promote Garlic Growth, this constitutes the first aspect of the invention.
The present invention also provides Bei Laisi bacillus to prepare the application in Garlic Growth promotor, including as activity
The Bei Laisi bacillus of ingredient and carrier, the growth promoter are the promotor for promoting garlic plant height.In addition, described
Biostimulation agent further includes compounding agent, and the compounding agent is gibberellin (GA), methyl α-naphthyl acetate (NAA), at least one in compound sodium nitrophenolate
Kind, this constitutes the second aspect of the invention.
Beneficial effects of the present invention: with Bei Laisi bacillus promote Garlic Growth, non agricultural chemical residuum, non-environmental-pollution,
Security threat is not caused to people and animals；With Garlic Growth promotor prepared by the fermentation liquid of Bei Laisi bacillus, preparation process letter
Single, at low cost, effect is good, easy to use.
Bei Laisi bacillus of the invention is Bacillus subtilis strain NDS11, document source: Li Le
The separation of the garlic endophyte such as small stream and the screening of antagonistic strain and identification [J] Jiangsu's agriculture science, 2018,46 (5): 97-
101. Bei Laisi bacillus subtilis strain NDS11 can plant big from Xuzhou City of Jiangsu Province Pei County
In garlic phosphorus stem by conventional method separation screening and obtain, the applicant can also be contacted and asked for.Applicant states from the application Shen
Please be from day in 20 years, the public can contact applicant and ask for the strain.
Detailed description of the invention
Fig. 1 is the 16SrRNA Phylogenetic Tree of bacterial strain 125 described in the embodiment of the present invention 1；
Fig. 2 is the photo that bacterial strain 125 described in the embodiment of the present invention 2 produces auxin (IAA) in vitro；Wherein, left figure
The photo for producing IAA for 125, right figure are LB blank control；
Fig. 3 is the photo that bacterial strain 125 promotes Garlic Growth in the greenhouse described in the embodiment of the present invention 3, wherein left
Figure is the control for not connecing 125 bacterium solution of bacterial strain, and right figure is the processing of 125 bacterium solution of inoculating strain.
Specific embodiment:
Embodiment 1: isolation and identification for strains
In the garlic phosphorus stem of Xuzhou City of Jiangsu Province Pei County acquisition health, endophyte is extracted by conventional method, and separate pure
Change, obtaining one plant of pure strain number is 125.Using LB liquid medium in 37 DEG C, the culture of 200rpm shaking flask for 24 hours, later with
Thalline were collected by centrifugation by 8000rpm, then by multistage microwave amplifier phage gene group DNA, finally uses bacterial universal primers 27F:5'-
AGAGTTTGATCCTGGCTCAG-3'(sequence 2) and 1492R:5'-AAGGAGGTGATCCAGCCGCA-3'(sequence 3) carry 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.It is soft using Mega5
Part phylogenetic tree construction is as shown in Figure 1, show that bacterial strain 125 should be a member in Bacillus subtilis.In NCBI and
EzBio Cloud compares sequencing result on website, and comparison result is as shown in table 1, the bacterial strain and Bei Laisi bacillus
The similitude of Bacillus subtilis strain NDS11 bacterial strain is 100%.
The sequence alignment result of 1 number of table, 125 bacterial strain
Embodiment 2: bacterial strain 125 generates the measurement for promoting plant growth substance in vitro
Bacterial strain 125 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 125 bacterial strains, if compared with the control, connecing bacterium and handling out
Existing pink then shows that 125 bacterial strains produce IAA (Fig. 2), indicates that 125 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 125 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 2 125 bacterial strain of table generation growth-promoting substance
Embodiment 3: bacterial strain 125 promotes Garlic Growth measure of merit
The 125 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 125 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 125, growing way is luxuriant, and
Blade is broad, shows that bacterial strain 125 is significant to the promotion growth result of garlic.
Due to the symbiosis of Bei Laisi bacillus and garlic, Bei Laisi bacillus is to have to the healthy growth of garlic
Benefit, therefore, the growth that can promote garlic on garlic is being sprayed from being separated after endophyte is enriched in garlic.Bei Laisi
Bacillus can mix with nonionic surfactant, such as tween, fatty amine, glycerine fatty acid esters, using water as medium, ultra micro
Thick flowable suspension is made in crushing, is sprayed on crop surface, adsorbable that different dispersions is formed in microparticle surfaces
System improves the distribution and infiltration of plant basal part of stem biological pesticide, increases operation rate.Secondly, can also be with inert filler and a certain amount of
Dispersing agent reach certain particle size in proportion after mixing well.These powdered pesticides can directly impose on plant stem foot
Portion is absorbed, and can also be added water-wet profit, dispersion, is spray applied.Powdered pesticide has the characteristics that storage and transportation is convenient, safe, packaging material
It is easily processed, 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>Bei Laisi bacillus is promoting the application in Garlic Growth
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1379
<212> DNA
<213>Bei Laisi bacillus (Bacillus velezensis)
<400> 1
aaaggttacc tcaccgactt cgggtgttac aaactctcgt ggtgtgacgg gcggtgtgta 60
caaggcccgg gaacgtattc accgcggcat gctgatccgc gattactagc gattccagct 120
tcacgcagtc gagttgcaga ctgcgatccg aactgagaac agatttgtgg gattggctta 180
acctcgcggt ttcgctgccc tttgttctgt ccattgtagc acgtgtgtag cccaggtcat 240
aaggggcatg atgatttgac gtcatcccca ccttcctccg gtttgtcacc ggcagtcacc 300
ttagagtgcc caactgaatg ctggcaacta agatcaaggg ttgcgctcgt tgcgggactt 360
aacccaacat ctcacgacac gagctgacga caaccatgca ccacctgtca ctctgccccc 420
gaaggggacg tcctatctct aggattgtca gaggatgtca agacctggta aggttcttcg 480
cgttgcttcg aattaaacca catgctccac cgcttgtgcg ggcccccgtc aattcctttg 540
agtttcagtc ttgcgaccgt actccccagg cggagtgctt aatgcgttag ctgcagcact 600
aaggggcgga aaccccctaa cacttagcac tcatcgttta cggcgtggac taccagggta 660
tctaatcctg ttcgctcccc acgctttcgc tcctcagcgt cagttacaga ccagagagtc 720
gccttcgcca ctggtgttcc tccacatctc tacgcatttc accgctacac gtggaattcc 780
actctcctct tctgcactca agttccccag tttccaatga ccctccccgg ttgagccggg 840
ggctttcaca tcagacttaa gaaaccgcct gcgagccctt tacgcccaat aattccggac 900
aacgcttgcc acctacgtat taccgcggct gctggcacgt agttagccgt ggctttctgg 960
ttaggtaccg tcaaggtgcc gccctatttg aacggcactt gttcttccct aacaacagag 1020
ctttacgatc cgaaaacctt catcactcac gcggcgttgc tccgtcagac tttcgtccat 1080
tgcggaagat tccctactgc tgcctcccgt aggagtctgg gccgtgtctc agtcccagtg 1140
tggccgatca ccctctcagg tcggctacgc atcgtcgcct tggtgagccg ttacctcacc 1200
aactagctaa tgcgccgcgg gtccatctgt aagtggtagc cgaagccacc ttttatgtct 1260
gaaccatgcg gttcaaacaa ccatccggta ttagccccgg tttcccggag ttatcccagt 1320
cttacaggca ggttacccac gtgttactca cccgtccgcc gctaacatca gggagcaag 1379
<210> 2
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
aaggaggtga tccagccgca 20
Claims (6)
1. Bei Laisi bacillus (Bacillus velezensis) is promoting the application in Garlic Growth.
2. Bei Laisi bacillus according to claim 1 is promoting the application in Garlic Growth, which is characterized in that it has
Body includes, and when garlic bulb grows to 6 leaf, the bacterium solution of the Bei Laisi bacillus is sprayed at garlic plant basal part of stem.
3. the Bei Laisi bacillus in claim 1 is preparing the application in Garlic Growth promotor.
4. Bei Laisi bacillus according to claim 3 exists preparing the application in Garlic Growth promotor, feature
In the Garlic Growth promotor is the promotor for increasing garlic plant height.
5. Bei Laisi bacillus according to claim 4 exists preparing the application in Garlic Growth promotor, feature
In the Garlic Growth promotor includes as the Bei Laisi bacillus of active constituent and carrier.
6. the application in Garlic Growth promotor is being prepared according to Bei Laisi bacillus as claimed in claim 3 to 5,
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 |
|---|---|---|---|
| CN201811559654.2A CN109566656A (en) | 2018-12-20 | 2018-12-20 | Bei Laisi bacillus is promoting the application in Garlic Growth |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811559654.2A CN109566656A (en) | 2018-12-20 | 2018-12-20 | Bei Laisi bacillus is promoting the application in Garlic Growth |
Publications (1)
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| CN109566656A true CN109566656A (en) | 2019-04-05 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811559654.2A Pending CN109566656A (en) | 2018-12-20 | 2018-12-20 | Bei Laisi bacillus is promoting the application in Garlic Growth |
Country Status (1)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111690576A (en) * | 2020-07-24 | 2020-09-22 | 江苏师范大学 | Bacillus belgii MMB-51 for producing biosurfactant and application thereof |
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| CN111690576A (en) * | 2020-07-24 | 2020-09-22 | 江苏师范大学 | Bacillus belgii MMB-51 for producing biosurfactant and application thereof |
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