CN110452832A - One plant of acid resistance bacillus amyloliquefaciens Kc-5 and its application - Google Patents
One plant of acid resistance bacillus amyloliquefaciens Kc-5 and its application Download PDFInfo
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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
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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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
Abstract
The present invention provides one plant of acid resistance bacillus amyloliquefaciens Kc-5 and its applications.Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Kc-5, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on April 29th, 2019, referred to as CGMCC, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and deposit number is CGMCC No.17654.Bacillus amyloliquefaciens Kc-5 of the invention has extensive pH Bioclimatic analysis (pH4-10) and preferable acid resistance；The bacterium antimicrobial spectrum is wide, all there is good antagonism to the various plants such as Fusarium oxysporum, banana blight bacteria, phytoph-thora capsici leonian, Botrytis cinerea and Cochliobolus sativus disease fungus, potting and field trial show that the bacterium has good disease prevention growth-promoting effect to the banana blight in acid soil.The bacterial strain adaptability is strong, and bacteriostatic activity is strong, can use in neutral and acid soil, the efficient prevention and control for fungal disease；The bacterium is also suitable for the improvement of acid soil.
Description
Technical field
The present invention relates to microorganisms technical fields.
Background technique
Fungal disease accounts for 70% of plant infectious diseases or more, is that crops and fruit tree yield and quality is caused to reduce
Main reason.The frequent use of chemical pesticide causes some disease fungus drug resistances sharply to enhance, cause environment and agricultural production
The pollution of product, and destroy microbial balance inside and outside plant and aggravate the eruption and prevalence of disease etc. on (remaining phoenix jade etc. 2005).Benefit
It is an ideal alternative route with the biological control that microorganism carries out fungal pathogens.Biological control noresidue is pollution-free, disease
Opportunistic pathogen will not generate resistance, and at low cost, increasing both production and income is conducive to the sustainable development of agricultural, has broad prospects.
Bacillus is most potential and commercial value one of biocontrol microorganisms, has has a broad antifungal spectrum, grows rapid, Yi Pei
It supports, the advantages that resistance is strong, biological safety is high and control efficiency stablizes, all the time to the study on prevention of plant disease
It is a hot and difficult issue.For many years people screened many Bacillus strains with Biocontrol Potential and develop at
Commercial preparation is applied to production practices.As american goods bacillus biocontrol bacterial strain QST713, GB03, MB1600 and
FZB24 etc., and the bent prevention and control for being used for plant disease such as peaceful of domestic bacillus commercial preparation sub- precious, hundred anti-, Mai Fengning, line
(leaf Jingjing etc. 2016).Although these biocontrol bacterial strains all have stronger antifungal activity, and have certain Field information effect,
But it is narrow that there are environmental suitabilities, usually there is certain region limitation (Neeraja et al.2010) in the application.Especially
The most suitable growth pH range of bacterium is in weak acid, neutrality to alkalinity.Many Biocontrol Bacillus are in weak acid and neutral soil to true
Fungus diseases have good prevention and control to act on, but to plant disease in acid soil then without control effect, this is often as these
Caused by bacillus can not colonize in acid soil.In recent years, people are also separated to some acid proof buds from animal intestinal tract
Spore bacillus is used as probiotics or feed addictive for improving animal and bird intestines environment, improves efficiency of feed utilization and animal
Immunity (Chen Yaqian etc. 2018；Open snow etc. 2018；2018) hair is fine to be waited.But there is presently no acid resistance bacillus to be used for plant
The report of disease biological and ecological methods to prevent plant disease, pests, and erosion and soil improvement limits bacillus to the application range of plant disease biological and ecological methods to prevent plant disease, pests, and erosion.
Bacillus amyloliquefaciens Kc-5 is the wide, broad-spectrum antifungal with ambient adaptability that we separate from acid soil
The bacillus of activity and biological and ecological methods to prevent plant disease, pests, and erosion application potential, by Molecular Identification, it is bacillus amyloliquefaciens (Bacillus
amyloliquefaciens).Research shows that bacillus amyloliquefaciens are a kind of with the active bacterium of broad-spectrum antibacterial, can generate
A variety of antibacterial substances, while it also has the function of promoting plant growth, has in environmental protection, planting industry, aquaculture and widely answers
With prospect (Guan Xiaohuan Jiang Hua 2012；Wang Jihuas etc. are 2017).But acid resistance bacillus and the microorganism prepared using it as material
There is not been reported for plant disease biological and ecological methods to prevent plant disease, pests, and erosion and soil improvement for preparation, microbial manure or soil conditioner.
Summary of the invention
The first object of the present invention is to provide bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Kc-5,
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 29th, 2019, referred to as
CGMCC, address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and deposit number is CGMCC No.17654.
The second object of the present invention is to provide a kind of microorganism formulation, and active constituent is bacillus amyloliquefaciens Kc-5.
The third object of the present invention is to provide bacillus amyloliquefaciens Kc-5 in preparing microbial manure and biological pesticide
Application.
The fourth object of the present invention be to provide bacillus amyloliquefaciens Kc-5 preparation prevention and treatment banana blight fertilizer or
Application in pesticide.
The fifth object of the present invention is to provide bacillus amyloliquefaciens Kc-5 and is preparing answering in banana growth-promoting agent or fertilizer
With.
What the present invention was protected is bacillus amyloliquefaciens Kc-5 and the microorganism formulation prepared with the bacterium, microorganism fertilizer
Material or soil conditioner and its application technology.
Bacillus amyloliquefaciens Kc-5 of the invention has extensive pH Bioclimatic analysis (pH4-10) and preferable acid resistance；
The bacterium antimicrobial spectrum is wide, to Fusarium oxysporum, banana blight bacteria, phytoph-thora capsici leonian, Botrytis cinerea and root rotof flax
The various plants such as bacterium disease fungus all has good antagonism, and potting and field trial show the bacterium in acid soil
Banana blight has good disease prevention growth-promoting effect.The bacterial strain adaptability is strong, and bacteriostatic activity is strong, can be in neutral and acid soil
It is used in earth, the efficient prevention and control for fungal disease；The bacterium is also suitable for the improvement of acid soil.
Detailed description of the invention
Fig. 1 is the pH areal map of bacillus amyloliquefaciens Kc-5 morphological feature and suitable growth.Wherein, A is bacterial strain Kc-5
Colonial morphology (left side) and Gram's staining (right side) on LB plate, B are growth feelings of the bacterial strain Kc-5 on different pH culture mediums
Condition.
Fig. 2 is antagonism figure of the bacillus amyloliquefaciens Kc-5 to various plants pathogen.
Fig. 3 is the results from pot experiment test figure of bacillus amyloliquefaciens Kc-5 preventing and controlling banana fusarium wilt.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.
PDA culture medium: potato dextrose agar, potato 200g filter to take filtrate, Portugal after boiling 30 minutes
Grape sugar 20g, agar 15-20g, with water constant volume to 1000ml, natural pH；115 DEG C of high-temperature sterilization 30min.
LB culture medium: tryptone 10g, yeast extract 5g, NaCl 5g, agar 15-20g (solid medium) use water
Constant volume is to 1000ml, with 1mol/L NaOH tune pH to 7.4,115 DEG C of high-temperature sterilization 30min.
Embodiment 1
Separation, identification and the biological characteristics of bacterial strain
One, the separation and purifying of bacterial strain
This experiment isolated several plants of bacteriums, process from the acid soil of Guangxi is as follows: 1g sample is added on equipped with small
In bead and the triangular flask of 100ml sterile saline, 28 DEG C of shaking table enrichment culture 30min or so.1m suspension is taken, using ladder
It spends diluted method and is diluted to 10-4, take stoste, 10-2Dilution and 10-4Dilution is cultivated at least two weeks after distinguishing coated plate.It grows
Bacterium colony is using plate streak in carrying out choosing single colonie after purification and being inoculated on LB plate on LB plate.It is protected using 20% glycerol
Gained purifying bacterial strain is deposited in -80 DEG C of refrigerators.
Two, the identification of bacterial strain
1, Molecular Identification
PCR amplification 16S rDNA uses universal primer 27F and 1492R, and (annealing temperature is about for 56 DEG C, target sequence
1500bp)。
Primer sequence is as follows: 27F:AGAGTTTGATCCTGGCTCAG-3；1492R:TACGGCTACCTTACGACTT.
Universal primer p-gyrA-f and p-gyrA-r (the annealing temperature that DNA amplification gyrase A subunit coding gene gyrA is used
Degree is 60 DEG C, and target sequence is about 1017bp).
Primer sequence is as follows: p-gyrA-f:CAGTCAGGAAATGCGTACGTCCTT；p-gyrA-r:
CAAGGTAATGCTCCAGGCATTGCT。
PCR response procedures are as follows: 95 DEG C of 5min；95 DEG C of 30s, 56 DEG C/60 DEG C 90s, 72 DEG C of 60s (33cycles)；72℃
10min；4℃.
The DNA fragmentation of amplification is carried out sequencing after purification, carries out sequence alignment by the Blastn program of NCBI.
Pass through 16S rDNA (SEQ ID NO.1) and DNA gyrase A subunit coding gene gyrA (SEQ ID NO.2) sequence analysis mirror
It is set to bacillus amyloliquefaciens (Bacillus amyloliquefaciens), is named as bacillus amyloliquefaciens Kc-5.
2, morphology and physio-biochemical characteristics
Morphologic observation is carried out after LB culture medium inoculated culture with the method for plate streaking；Pass through Gram stain analysis
Thallus is Gram-negative bacteria or positive bacteria, and observes thallus and the form of brood cell simultaneously.
The water stain shape of early stage bacterium colony, middle and later periods bacterium colony milky, there is crateriform protuberance, and edge is irregular.Bacterial strain Kc-5 is
Gram-positive bacteria, rod-shaped, endogenous spore, atrichia.The pH of suitable growth is 4-10.Suitable growth temperature is 28-38 DEG C.
Three, the biological characteristics of bacterial strain
The pH and temperature range of the biological characteristics Main Analysis suitable growth of bacterial strain.
With the LB liquid medium of the buffer difference pH (pH3-10) of different buffer systems, it is inoculated with starch gemma bar
Bacterium Kc-5 in equipped with different pH fluid nutrient mediums test tube in, 30 DEG C of shaking table cultures 24 as a child observed the variation of biomass with
Analyze the pH range of bacterial strain Kc-5 suitable growth.
Bacillus amyloliquefaciens Kc-5 water stain shape of early stage bacterium colony on LB culture medium, middle and later periods bacterium colony milky, there is volcano
Mouth shape protuberance, edge are irregular (Fig. 1).Physiological and biochemical test shows that bacterial strain Kc-5 is gram-positive bacteria, rod-shaped, inside sprouts
Spore.The pH of suitable growth is 4-10 (Fig. 1).Suitable growth temperature is 28-38 DEG C.
Four, the preservation of bacterial strain
It is isolated from the acid resistance bacillus amyloliquefaciens Kc-5 of acid soil, it is micro- that China is preserved on April 29th, 2019
Biological inoculum preservation administration committee common micro-organisms center, referred to as CGMCC, address are BeiChen West Road, Chaoyang District, BeiJing City 1
Number institute 3, deposit number are CGMCC No.17654.
Embodiment 2
The antifungal activity of acid resistance bacillus amyloliquefaciens Kc-5 is analyzed
1, bacillus amyloliquefaciens Kc-5 culture and collection
Using method of scoring inoculation Kc-5 on LB plate, after growing single colonie, picking single colonie is inoculated in liquid LB training
Support base in 30 DEG C culture 24 hours after it is spare.
2, the culture of pathogen
It is inoculated with banana blight bacteria, cucumber fusarium axysporum, the flat navel child spore bacterium of Root Rot of Wheat, P. capsici, vegetable ash
In on PDA plate, 28 DEG C of cultures use mildew bacterium after a week.
3, plate face-off experimental result
Inoculation Bacteria culturing object fungus block is being inoculated with solution starch apart from pathogen 2.0cm or so in PDA plate center respectively
Bacillus Kc-5 bacteria suspension, 28 DEG C are cultivated 3-5 days, observe growth of pathogenic bacteria situation.
The acid resistance bacillus amyloliquefaciens Kc-5 for being isolated from acid soil has the antifungal activity of wide spectrum.
Plate face-off experiment display, Kc-5 is to Fusarium oxysporum Cuba specialized form (Fusariun oxysporun
F.sp.cubense), Fusarium oxysporum cucumber specialized form (F.oxysporum f.sp.cucumerinum), Phytophthora capsici
(Phytophthora capsici), the pathogen of Botrytis cinerea (Botrytis cinereal), flat navel child spore bacterium (Bipolaris
) etc. sorokiniana various plants disease fungus all has good antagonism.
Embodiment 3
The preparation of acid resistance bacillus amyloliquefaciens Kc-5 microbial inoculum
1. the preparation of seed liquor
Picking grows 24 hours Kc-5 single colonies on LB plate, is seeded to containing 150ml LB liquid medium
In 500ml triangular flask, 37 DEG C, 180rpm is cultivated 23 hours.Then it is forwarded to according to 3-5% ratio and new contains 150ml liquid
In the l triangular flask of LB culture medium, then training 6 hours is shaken, is used as seed liquor.
2. the preparation of liquid fermentation microbial bacterial agent
In 50L fermentor contain 30L culture medium: yeast powder 3g/l, peptone 5g/L, beef extract 2g/L, sodium chloride 2g/L,
MnSO40.005g/L, K2HPO43g/L, MgSO40.02g/L.PH: keeping pH is 7.0, and fermentation process is adjusted with ammonium hydroxide.
DO: dissolved oxygen is associated with revolving speed, keeps about 30%.
Tank pressure: 0.04Mpa.
Stream plus glucose: when residual sugar content reduces, (about 4-5 hours) starts to flow and add, and substantially remains in 5g/L or more.
37 DEG C of fermentations to end in 40 hours is fermented, microscopy gemma rate > 90%.
After liquid fermentation, fermentation liquid is centrifuged, collects thallus.
Kieselguhr adsorption thallus is added according to the 3% of fermentating liquid volume, mixture is dried by fluidized bed, and temperature control exists
60 DEG C hereinafter, moisture control crosses 80 meshes, obtains the microbial bacterial agent prepared by liquid fermentation mode within 10%.
3. the preparation of solid fermentation microbial bacterial agent
Using corn flour, wheat bran, dregs of beans as base stock, a certain proportion of K is added2HPO4、MnSO4、MgSO4, material-water ratio 1:
0.8.Seed liquor is inoculated with according to 1% ratio.37 DEG C are cultivated 4-5 days, and spore concentration can achieve 1 × 1010Hundred million/g, as solid
Microbial inoculum uses.
Embodiment 4
The application of acid resistance bacillus amyloliquefaciens Kc-5
One, acid resistance bacillus amyloliquefaciens Kc-5 tests the potting prevention and control of banana blight
1, experimental material and method
1) potted plant experiment is using clay and nutrient matrix composite soil, pH5.5 or so, every basin dress soil 1kg.Banana seedlings kind is
Osmanthus any of several broadleaf plants No. 6, plant height 30cm or so.
2) No. 4 biological strains of banana blight bacteria (F.oxysporun f.sp.cubense) are cultivated in PDA culture medium
To spore is produced, being then made into concentration with physiological saline is 107The spore suspension of a/ml is spare.
3) prepare bacillus amyloliquefaciens Kc-5 microbial inoculum according to embodiment 3, being made into concentration with physiological saline is 108CFU/ml
Spore suspension it is spare.
4) 6 basins of each processing, every basin plant 2 plants of banana seedlings.Other than blank control, remaining is handled banana blight bacteria
No. 4 biological strain bacteria suspensions are infiltrated in soil, make its final concentration of 105A/g.
5) it when transplanting, is planted in the soil after banana seedlings are handled 10-15 minutes with biocontrol microorganisms root dipping.It is again that biological and ecological methods to prevent plant disease, pests, and erosion bacterium solution is equal
It is even drench into containing bacterium soil in, make its final concentration of 107CFU/g.Blank control is set in experiment and does not apply the morbidity control of biocontrol microorganisms.
The banana seedlings of all processing are in plastic greenhouse culture.External symptom incidence is investigated every 7d, calculates disease index.Wait compare
Morbidity is obvious, splits banana seedlings basal part of stem investigation internal sympton incidence, calculates disease index.
The Disease investigation grade scale of (2003) such as severity Scaling standard reference Saravanan, to the outer of morbidity banana seedlings
Portion's symptom and bulb symptom, carry out sick grade statistics, and grade scale is shown in Table 1.
The grade scale of table 1 external symptom and bulb symptom
Routinely formula calculates disease index:
Disease index=feelings (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × superlative degree value) × advanced value
Opposite control efficiency (%)=(control disease index-processing disease index)/control disease index × shines the state of an illness.
2, experimental result
After transplanting 3 months, morbidity control group morbidity is serious, disease index 79, death rate 58.3%.And acid resistance solution starch
Bacillus Kc-5 processing group disease index 38, death rate is only 25%, reaches 51% or more to the preventive effect of banana blight bacteria
(Fig. 3).Kc-5 also has certain growth-promoting functions simultaneously, and the plant height and leaf area of the plant of Kc-5 microbial inoculum processing are significantly better than that
Control group (Fig. 3).
The acid resistance bacillus amyloliquefaciens Kc-5 of acid soil is isolated to banana blight (F.oxysporun
F.sp.cubense control effect).In the pot experiment prevented and treated banana blight, potting soil pH is 6 or so, banana
Kc-5 microbial inoculum root dipping and root irrigation are used when seedling replanting, can significantly mitigate the occurring degree of banana blight.And Kc-5
Microbial inoculum also has certain growth-promoting functions, and the plant height and leaf area of the plant of Kc-5 microbial inoculum processing are significantly better than that control group.
Two, field trial of the acid resistance bacillus amyloliquefaciens Kc-5 to banana blight
1, experimental material and method
1) test site and environmental condition: the experiment is located at Nanning Nanjing Zhen Hetan Luo Zhen.Two blocks of experimental plot soil
PH belongs to Acid soil between 4.5-5.0, this two pieces of field plantation bananas many years, banana blight disease incidence 50%
More than, have accumulated a large amount of banana blight bacteria.
2) plot area is arranged with processing: each field setting compares 0.5 mu of (application microbial inoculum), microbial inoculum Kc-5 processing 0.5
60 plants of banana (0.5 mu) is planted in mu, each processing.
3) microbial inoculum method of administration: testing preceding soil and turned over, and testing banana seedlings kind used is to be easy to infect banana
The kind Williams of wilt disease.Setting control (not applying microbial inoculum) and microbial inoculum Kc-5 processing when transplanting seedlings.Microbial inoculum processing sample into
Row cave is applied, and the part soil that every cave is dug out is mixed with 10g microbial inoculum (10,000,000,000/g), and then seedling is planted in backfill.In banana root system after 6 months
Surrounding adds a microbial inoculum (every plant of 10g microbial inoculum) by cave applying method.
4) result counts: incidence investigation and output statistics are carried out after 10 months.
2 experimental results
The field test results of 2 bacillus amyloliquefaciens Kc-5 preventing and controlling banana fusarium wilt of table
Investigation result shows (table 2) that Nanjing town field compares disease incidence 51.72%, and per mu yield is only 1737 kilograms；Microbial inoculum
It is only 2.78% that Kc-5, which handles disease incidence, preventive effect 94.62%, and per mu yield is 3500 kilograms, relatively control volume increase 101%.The town Tan Luo
Field control disease incidence is up to 64.79%, and per mu yield is only 1267 kilograms；It is only 27.59% that microbial inoculum Kc-5, which handles disease incidence, preventive effect
It is 57.42%, per mu yield is 2607 kilograms, relatively control volume increase 106%.It is shown by two places test result, for banana blight
Disease incidence 50% and banana field below, microbial inoculum Kc-5 can reach 94% or more preventive effect, and 100% or more volume increase is a kind of
It can colonize and play a role in acid soil.
Sequence table
<110>Institute of Microorganism, Academia Sinica
<120>one plants of acid resistance bacillus amyloliquefaciens Kc-5 and its applications
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1378
<212> DNA
<213>bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
<400> 1
gagcttgctc cctgatgtta gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta 60
agactgggat aactccggga aaccggggct aataccggat ggttgtctga accgcatggt 120
tcagacataa aaggtggctt cggctaccac ttacagatgg acccgcggcg cattagctag 180
ttggtgaggt aacggctcac caaggcgacg atgcgtagcc gacctgagag ggtgatcggc 240
cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtagg gaatcttccg 300
caatggacga aagtctgacg gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa 360
agctctgttg ttagggaaga acaagtgccg ttcaaatagg gcggcacctt gacggtacct 420
aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg 480
ttgtccggaa ttattgggcg taaagggctc gcaggcggtt tcttaagtct gatgtgaaag 540
cccccggctc aaccggggag ggtcattgga aactggggaa cttgagtgca gaagaggaga 600
gtggaattcc acgtgtagcg gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag 660
gcgactctct ggtctgtaac tgacgctgag gagcgaaagc gtggggagcg aacaggatta 720
gataccctgg tagtccacgc cgtaaacgat gagtgctaag tgttaggggg tttccgcccc 780
ttagtgctgc agctaacgca ttaagcactc cgcctgggga gtacggtcgc aagactgaaa 840
ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa 900
cgcgaagaac cttaccaggt cttgacatcc tctgacaatc ctagagatag gacgtcccct 960
tcgggggcag agtgacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg 1020
ttaagtcccg caacgagcgc aacccttgat cttagttgcc agcattcagt tgggcactct 1080
aaggtgactg ccggtgacaa accggaggaa ggtggggatg acgtcaaatc atcatgcccc 1140
ttatgacctg ggctacacac gtgctacaat ggacagaaca aagggcagcg aaaccgcgag 1200
gttaagccaa tcccacaaat ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt 1260
gaagctggaa tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct 1320
tgtacacacc gcccgtcaca ccacgagagt ttgtaacacc cgaagtcggt gaggtaac 1378
<210> 2
<211> 1017
<212> DNA
<213>bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
<400> 2
tcagtcagga aaagcgtacg tccttcctgg actatgcaat gagcgttatc gtatcccggg 60
cgcttccgga tgtgcgtgac ggtctgaagc cggttcacag gcggattttg tacgcaatga 120
atgatttagg catgaccagt gacaaaccat ataaaaaatc tgcccgtatc gtcggtgaag 180
ttatcggtaa gtaccacccg cacggtgact cagcggttta cgaatcaatg gtcagaatgg 240
cgcaggattt taactaccgc tacatgcttg ttgacggaca cggcaacttc ggttcggttg 300
acggcgactc agcggccgcg atgcgttaca cagaagcgag aatgtcaaaa atcgcaatgg 360
aaatcctccg ggacattacg aaagatacga ttgattatca agataactat gacggcgcag 420
aaagagaacc tgtcgtcatg ccttcgagat ttccgaatct gctcgtaaac ggagctgccg 480
gtattgcggt cggaatggcg acaaatattc ctccgcatca gcttggggaa gtcattgaag 540
gcgtgcttgc cgtaagtgag aatcctgaga ttacaaacca ggagctgatg gaatacatcc 600
cgggcccgga ttttccgact gcaggtcaga ttttgggccg gagcggcatc cgcaaggcat 660
atgaatccgg acggggatcc attacgatcc gggctaaggc tgaaatcgaa gagacatcat 720
cgggaaaaga aagaattatt gtcacagaac ttccttatca ggtgaacaaa gcgagattaa 780
ttgaaaaaat cgcagatctt gtccgggaca aaaaaatcga aggaattacc gatctgcgtg 840
acgaatccga ccgtaacgga atgagaatcg tcattgagat ccgccgtgac gccaatgctc 900
acgtcatttt gaataacctg tacaaacaaa cggccctgca gacgtctttc ggaatcaacc 960
tgctggcgct cgttgacgga cagccgaagg tactaagcct gaagcaatgc ctggagc 1017
Claims (7)
1. bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Kc-5 CGMCC No.17654.
2. a kind of microorganism formulation, active constituent bacillus amyloliquefaciens Kc-5.
3. bacillus amyloliquefaciens Kc-5 is preparing the application in microbial manure and pesticide.
4. application of the bacillus amyloliquefaciens Kc-5 in preparation prevention and treatment banana blight fertilizer or pesticide.
5. bacillus amyloliquefaciens Kc-5 is preparing the application in banana growth-promoting agent or fertilizer.
6. application of the bacillus amyloliquefaciens Kc-5 in preparation plant pathogenic fungi preparation, fertilizer, pesticide.
7. application according to claim 6, which is characterized in that the plant pathogenic fungi includes: to Fusarium oxysporum Cuba
Specialized form, Fusarium oxysporum cucumber specialized form, the pathogen of Botrytis cinerea, flat navel child spore bacterium and P. capsici etc..
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