CN107400638A - A kind of suitable raw bacillus amyloliquefaciens EZ10 01 of South Pole low temperature and its application - Google Patents
A kind of suitable raw bacillus amyloliquefaciens EZ10 01 of South Pole low temperature and its application Download PDFInfo
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Abstract
The invention discloses a kind of suitable raw bacillus amyloliquefaciens EZ10 01 of South Pole low temperature and its application.Bacillus amyloliquefaciens（Bacillus amyloliquefaciens）EZ10 01, on December 26th, 2016 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, preserving number is CGMCC No.13495.The bacterial strain can be that a kind of low temperature fits raw bacterium in 10 DEG C of growths.EZ10 01 has preferable prevention effect to corn bacterial brown rot, and its biocontrol effect has good application prospect up to 55.3%.
Description
Technical field
The invention belongs to biotechnology and agricultural disease Prevention Technique field, is related to a kind of suitable raw solution starch bud of South Pole low temperature
Spore bacillus EZ10-01 and its application.
Background technology
It is a kind of caused by the general bacterium of pineapple (Pantoea ananatis) in the regional large-scale outbreak such as Hebei in recent years
Corn bacterial brown rot, Symptoms are the withered curling of blade tip, and vein produces yellow scab, yellowish-brown dry rot at stalk.It is long
People are paid close attention to corn bacterial disease less since phase, and due to lacking effective preventions, corn bacterial disease is to agriculture
The loss that industry production is brought is often extremely serious.
China is Maize Production and consumption big country, and corn seeding area and yield all occupy the second in the world.In recent years, by
In the expansion of corn seeding area, weather conditions change, the popularization of new varieties and the change of cropping system, the generation of maize diseases
It is in rising trend, bring serious harm to agricultural production and food security.Hebei is the big province of corn planting, no matter growing surface
Product or yield Jun Ju North Chinas are the first, and maize diseases are the main reason for influenceing Hebei Province corn yield and quality.
The generations for using chemical pesticide control field diseases in the past more, but it is outstanding day by day the drawbacks of this kind of controlling mode：One
Aspect easily makes pathogen develop immunity to drugs, and " two subtract " policy on the other hand more vigorously advocated with country is disagreed.At present, very
More people start to find answer from the angle of biological and ecological methods to prevent plant disease, pests, and erosion, and have had many biological pesticides to be applied in production practices, have taken
Obtained good economic benefit and ecological benefits.
Rhizosphere microorganism species is various, wherein, there are many bacteriums to promote plant growth, controlling plant diseases, increase
Crop yield etc., this quasi-microorganism are commonly referred to as plant growth-promoting rhizobacteria (Plant Growth-Promoting
Rhizobacteria, abbreviation PGPR).They have good mutualism relation between plant, can not only produce plant and lure
Impedance characteristic of disease, and the yield and quality of crop can be improved.So plant growth-promoting rhizobacteria have in control of plant disease it is critically important
Application value.
The content of the invention
The present invention provides a kind of bacillus amyloliquefaciens, targetedly solves to ask present in current control of plant disease
Topic.The bacterium is that low temperature fits raw bacterium, has fungistatic effect to pathogens such as corn bacterial brown rot germs, has potential biological and ecological methods to prevent plant disease, pests, and erosion to be worth.
It is another object of the present invention to propose the application of the public bacillus amyloliquefaciens.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 of the present invention, in 2016
On December 26, in is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, preserving number CGMCC
No.13495。
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) the EZ10-01CGMCC No.13495,
It is Biocontrol Bacillus isolated in a kind of soil from antarctic penguin aggregation ground.Dilution applies flat board and obtained after 80 DEG C of water-baths
.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) the EZ10-01CGMCC No.13495 are
Gram-positive bacterium, it is shaft-like.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) the EZ10-01CGMCC No.13495's
16S rRNA partial nucleotide has the sequence as shown in sequence table SEQ ID NO.1.The bacillus amyloliquefaciens
(Bacillus amyloliquefaciens) EZ10-01CGMCC No.13495 gyrB partial nucleotide has such as sequence
Sequence shown in table SEQ ID NO.2.Bacterial strain of the present invention, reference《Bacillus》(Cai Miaoying etc. translates agriculturals and published
Society, 1988) and《Microbial taxonomy》The retrieval table of (Zhang Jizhong Fudan University in Shanghais publishing house, 1995) is retrieved,
It is identified on Physiology and biochemistry, and 16S rRNA and gyrB gene order results, it is Xie Dian through Bacillus subtilis
Afnyloliquefaciens (Bacillus amyloliquefaciens), are named as EZ10-01.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) the EZ10-01CGMCC No.13495 energy
It is that a kind of low temperature fits raw bacterium in 10 DEG C of growths.
A kind of biocontrol agent prepared by described bacillus amyloliquefaciens EZ10-01.
Described biocontrol agent is preferably prepared by the following method to obtain:By EZ10-01 inoculations on LB culture mediums,
It is incubated overnight, then the EZ10-01 bacterial strains of activation is transferred in Landy culture mediums and carry out shake flask fermentation, temperature is 30 DEG C, is shaken
Bed rotating speed is 200rpm, and fermentation time 35-40 hours to bacterium amount reach 2.46 × 109CFU/mL；Wherein Landy culture mediums form
For：20g glucose, 5g Pidolidones, 0.5g MgSO4, 0.5g KCl, 1g KH2PO4, 0.15mg Fe2SO4·6H2O,
5.0mg MnSO4·H2O, 0.16mg CuSO4·5H2O, pH7.0,1000mL distilled water.
Described bacillus amyloliquefaciens EZ10-01 is suppressing corn bacterial brown rot germ, rice leaf spot bacteria, water
Application in rice Xanthomonas campestris PV.oryzicola, fusarium graminearum and/or corn wheel branch Fusariumsp.
Application process is that fermented and cultured EZ10-01 obtains zymotic fluid, and rear foliage-spray is diluted to zymotic fluid.
Described bacillus amyloliquefaciens EZ10-01 is preparing preventing and treating corn bacterial brown rot germ, bacterial blight of rice
Bacterium, xanthomonas oryzae pv. oryzicola, fusarium graminearum and/or corn wheel branch Fusariumsp biocontrol agent in application.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention, it is
It is a kind of to corn bacterial brown rot germ, rice leaf spot bacteria, xanthomonas oryzae pv. oryzicola, fusarium graminearum and/or
Corn wheel branch Fusariumsp has the bacillus of good preventive effect, is that a kind of low temperature fits raw bacterium, environmentally safe, potentially should have
With value.
Brief description of the drawings
Fig. 1 EZ10-01 growing states
A is EZ15-07, and B is control strain FZB42, C EZ10-01
Fungistatic effects of Fig. 2 EZ10-01 to corn bacterial brown rot germ
Field efficacies of Fig. 3 EZ10-01 to corn bacterial brown rot
A is CK, B EZ10-01
Fungistatic effects of Fig. 4 EZ10-01 to rice leaf spot bacteria
Fungistatic effect of Fig. 5 EZ10-01 bacterial strains to xanthomonas oryzae pv. oryzicola
Fungistatic effects of Fig. 6 EZ10-01 to fusarium graminearum
Fungistatic effects of Fig. 7 EZ10-01 to corn wheel branch Fusariumsp
Biomaterial preservation information
EZ10-01, Classification And Nomenclature is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), in being preserved in
State's Microbiological Culture Collection administration committee common micro-organisms center, preservation date are on December 26th, 2016, and preservation address is
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.13495.
Embodiment
The present invention is further illustrated with reference to embodiment.
Embodiment 1：
The sieve of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention
Choosing.
Pedotheque for separation is all from antarctic penguin settlement, and soil sample is collected in away from the 10-15cm of ground.Weigh
5g soil samples are shaken 30 minutes in 45mL physiological saline on shaking table, and standing makes clarification.Aspirate supernatant is dilute with sterilized water respectively
Release 10-1、10-2、10-3、10-4、10-5Five concentration gradients, 80 DEG C of water-bath 10min.Draw 200 μ L and apply LB flat boards, 37 DEG C overnight
Culture.After bacterium colony is grown on flat board, picking single bacterium colony, numbering preserves after purification for line.
Embodiment 2：
Bacillus amyloliquefaciens Bacillus amyloliquefaciens EZ10-01 bacterial strains provided by the invention resist
Low temperature screening process.
Specific implementation step：
Culture medium：
1C culture mediums
| Composition | Concentration | 1×1C | 5×1C |
| Peptone | 0.7 | 4.2g | 21g |
| Sodium chloride | 0.5 | 3g | 15g |
| Soy meal | 0.3 | 1.8g | 9g |
| Cumulative volume | Add water to 600mL | Add water to 600mL |
Soil extract
Whole culture medium
| Composition | Final concentration | Volume (ml) |
| 5×1C | 1× | 50 |
| 20% glucose | 0.1% | 1.25 |
| Soil extract | 10% | 25 |
| Water | 173.75 |
By inoculation into aforesaid liquid culture medium, 37 DEG C of overnight incubations, 5 μ L bacterium solution points are taken to apart from solid medium
Plate center 2.5cm right-angled intersection point, each 5 repetitions of bacterial strain, it is individually positioned in 4 DEG C, 10 DEG C, 14 DEG C, 18 DEG C, 30 DEG C of trainings
Support and cultivated in case, record strain growth situation daily, record 15 days.The bacterial strain can grow in the environment of temperature≤10 DEG C, I
Will at this temperature, the bacterial strain that bacterium colony can be shown in one week is designated as low-temperature epitaxy bacterial strain.Further testing the bacterial strain can be with
Grown under conditions of 10 DEG C, and after flat board culture 7d, will show the region of bacterium colony together with culture medium in the lump from flat on flat board
Peeled off on plate, be placed in centrifuge tube and add 1mL sterilized waters concussion 10min.Supernatant dilution is taken to apply plate count after concussion.This
When, one plant of bacterium EZ10-01 is screened, it is 3.88 × 10 that bacterium amount, which increases,7CFU/mL, it is 1.92 times of initial bacterium amount.Bacterium amount
Growth rate is 1.8 times of control strain FZB42.Growing state is as shown in Figure 1.
Embodiment 3：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention
The PCR amplifications of 16S rRNA genes and sequencing.
By the inoculation in LB liquid medium, 180rpm, 37 DEG C of concussion and cultivates 10 hours, thalline is collected by centrifugation, hangs
Lysozyme and SDS broken walls are added after floating, genomic DNA is extracted using phenol-chloroform method, with forward primer 27F (5 '-
AGAGTTTGATCMTGGCTCAG-3 ' (SEQ ID NO.3)) and reverse primer 1492R (5 '-GGYTACCTTGTTACGACTT-
3 ' (SEQ ID NO.4)), 16S rRNA genes enter performing PCR amplification, are carried out sending sequencing to be surveyed after PCR primer recovery
Sequence.PCR conditions are：95 DEG C, 4min；94 DEG C, 1min；50 DEG C, 1min；72 DEG C, 2min；34cycle；72 DEG C, 10min, 4 DEG C
Preserve.16S rRNA gene order length is about 1400bp, in GenBank with Bacillus amyloliquefaciens
Lx-11 similitudes are 99%.Its result is as shown in sequence table SEQ ID NO.1.
Embodiment 4：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention
The PCR amplifications of gyrB genes and sequencing.
By the inoculation in LB liquid medium, 180rpm, 37 DEG C of concussion and cultivates 10 hours, thalline is collected by centrifugation, hangs
Lysozyme and SDS broken walls are added after floating, genomic DNA is extracted using phenol-chloroform method, with forward primer UP1f (5 '-
GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3 ' (SEQ ID NO.5)) and reverse primer UP2r (5 '-
AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3 ' (SEQ ID NO.6)), gyrB genes are carried out
PCR is expanded, and is carried out sending sequencing to be sequenced after PCR primer recovery.PCR conditions are：95 DEG C, 4min；94 DEG C, 1min；
62 DEG C, 1min；72 DEG C, 2min；34cycle；72 DEG C, 10min, 4 DEG C preservations.GyrB gene order length is about 1200bp,
In GenBank with Bacillus amyloliquefaciens CC178 similitudes be 96%.Its result such as sequence table SEQ ID
Shown in NO.2.
Reference《Bacillus》(Cai Miaoying etc. translate agriculturals publishing house, 1988) and《Microbial taxonomy》(Zhang Ji
Loyal Fudan University in Shanghais publishing house, 1995) retrieval table is retrieved, and it is identified on Physiology and biochemistry, and 16S
RRNA and gyrB gene order results, bacterium EZ10-01 is accredited as bacillus amyloliquefaciens (Bacillus through polyphase sort
amyloliquefaciens)。
Embodiment 5：
The hair of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention
Ferment culture.
By EZ10-01 inoculations on LB culture mediums, it is incubated overnight, is then transferred to the EZ10-01 bacterial strains of activation
Shake flask fermentation is carried out in Landy culture mediums, temperature is 30 DEG C, shaking speed 200rpm, and fermentation time is 38 hours.Wherein
Landy fermentation mediums form：20g glucose, 5g L-glutamic acid, 0.5g MgSO4, 0.5g KCl, 1g
KH2PO4, 0.15mg Fe2SO4·6H2O, 5.0mg MnSO4·H2O, 0.16mg CuSO4·5H2O, pH7.0,1000mL
distilled water.Bacterium amount reaches 2.46 × 10 after shake flask fermentation 38h9CFU/mL。
Embodiment 6：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention are to jade
The bacteriostasis of rice Bacterial brown rot bacterium.
Using flat board bacteriostatic method, draw 5mL Liquid Cultures 18h corn bacterial brown rot pathogen bacterium solution in cool to
Plate drying is down flat after being shaken up in 40 DEG C or so of LB culture mediums.The aseptic filter paper piece in above-mentioned flat board, draw Liquid Culture 18h's
The μ L of bacillus bacterium solution 5 are added dropwise on filter paper, 37 DEG C of incubated 20h.EZ10-01 is observed to corn bacterial brown rot
The fungistatic effect of bacterium, average diameter of inhibition zone 12.7mm, as shown in Figure 2.
Embodiment 7：
Corn bacterial brown rot can be suppressed by zymotic fluid, detailed process is：Bacterial strain EZ10-01 is fermented,
Zymotic fluid carries out foliage-spray processing after diluting 100 times.Foliage-spray is carried out in the pustulation period of corn, its biocontrol effect is reachable
55.3%.Fermentation culture conditions are：By EZ10-01 inoculations on LB culture mediums, it is incubated overnight, then by activation
EZ10-01 bacterial strains, which are transferred in Landy culture mediums, carries out shake flask fermentation, and temperature is 30 DEG C, shaking speed 200rpm, during fermentation
Between be 38 hours.Field efficacy is as shown in Figure 3.
Embodiment 8：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention are to water
The bacteriostasis of rice bacterial leaf spot pathogenic bacteria.
Using flat board bacteriostatic method, added in 150mL NA culture mediums and cultivated rice leaf spot bacteria 5mL, NA is put down
Plate, each flat board put the filter paper of four sterilizings, and the bacillus bacteria suspension for then drawing 5 μ L is dropped on filter paper, 28 DEG C of trainings
Result is observed after supporting 3 days.It is repeated 3 times, fungistatic effects of the observation bacterial strain EZ10-01 to rice leaf spot bacteria.Inhibition zone is averaged
A diameter of 21.3mm, as shown in Figure 4.
Embodiment 9：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention are to water
The bacteriostasis of rice Xanthomonas campestris PV.oryzicola.
Using flat board bacteriostatic method, added in 150mL NA culture mediums and cultivated xanthomonas oryzae pv. oryzicola 5mL, fallen
NA flat boards, each flat board put the filter paper of four sterilizings, and the bacillus bacteria suspension for then drawing 5 μ L is dropped on filter paper, and 28
DEG C culture 3 days after observe result.It is repeated 3 times, fungistatic effects of the observation bacterial strain EZ10-01 to xanthomonas oryzae pv. oryzicola.Suppression
Bacterium circle average diameter is 17.6mm, as shown in Figure 5.
Embodiment 10：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention are to small
The bacteriostasis of wheat gibberellic hypha.
Using flat board face-off method, good PDA plate, diameter 1cm fusarium graminearum fungus block is placed in flat board center,
Symmetrically placed four filter papers at its surrounding distance 2cm, 5 μ L fermentation of bacillus liquid is added dropwise on filter paper, in triplicate,
Cultivated 3 days in 25 DEG C of illumination boxs, EZ10-01 is to the fungistatic effect of fusarium graminearum, average diameter of inhibition zone for observation
For 20mm, as shown in Figure 6.
Embodiment 11：
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01 bacterial strains provided by the invention are to jade
The bacteriostasis of rice wheel branch Fusariumsp.
Using flat board face-off method, good PDA plate, diameter 1cm corn wheel branch Fusariumsp fungus block is placed in flat board center,
Symmetrically placed four filter papers at its surrounding distance 2cm, 5 μ L fermentation of bacillus liquid is added dropwise on filter paper, repeats three
It is secondary, cultivated 3 days in 25 DEG C of illumination boxs, observe fungistatic effects of the EZ10-01 to corn wheel branch Fusariumsp, inhibition zone is average
A diameter of 37mm, as shown in Figure 7.
<110>Agricultural University Of Nanjing
<120>A kind of suitable raw bacillus amyloliquefaciens EZ10-01 of South Pole low temperature and its application
<160> 2
<210> 1
<211> 1451
<212> DNA
<213>Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01
<220>
<223> 16S rRNA
<400> 1
agaggcgggg tgctataatg caagtcgagc ggacagatgg gagcttgctc cctgatgtta 60
gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga 120
aaccggggct aataccggat ggttgtttga accgcatggt tcagacataa aaggtggctt 180
cggctaccac ttacagatgg acccgcggcg cattagctag ttggtgaggt aacggctcac 240
caaggcgacg atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg 300
gcccagactc ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg 360
gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa agctctgttg ttagggaaga 420
acaagtgccg ttcaaatagg gcggcacctt gacggtacct aaccagaaag ccacggctaa 480
ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540
taaagggctc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600
ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca 840
ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgacaatc ctagagatag gacgtcccct tcgggggcag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140
accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200
gtgctacaat ggacagaaca aagggcagcg aaaccgcgag gttaagccaa tcccacaaat 1260
ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320
tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380
ccacgagagt ttgtaacacc cgaagtcggt gaggtaacct tttaggagcc agccgccgaa 1440
ggtcacagat t 1451
<210> 2
<211> 1193
<212> DNA
<213>Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01
<220>
<223>GyrB genes
<400> 2
gggcacatga cgctagatta cctgctggga tacgcgcttt ttcaagatta aaatcttctc 60
cgattcctgt tccgagggcc gtgatcattg atctgacctc attgtttgag agaattttat 120
caagtctggc tttctcaacg ttcagaatct taccgcgcag cggcagaatg gcttggaaat 180
gacggtcccg tccctgtttc gctgatccgc ccgcagagtc accctctacg atatacagct 240
cggaaatgct cggatcttta gaagaacagt ccgccagttt gcccggcaga ttggaaatct 300
caagcgcact tttgcggcgg gtcaattccc gcgctttttt cgctgccatc cgcgctcttg 360
cggccattaa acctttttca acgattttgc gggctgagtc cggattttca agaaggaatg 420
tttccagcgc agaagaaaac agcgtatcag tgatcgttct cgcttcggag ttgccgagct 480
tcgttttcgt ctgcccttcg aattgcggat cagggtgctt aattgaaata atggcagtca 540
gcccttctct cacatcatcc ccgcttaaat tcggatcatt ttctttgaaa atcccttttc 600
ttcttgcata gtcgtttatg acacgggtca gaccggtttt aaatccggct tcgtgcgtcc 660
cgccttcgta tgtgttgatg ttatttgtga aagaataaat gttgcttgta tagctgtcgt 720
tgtattgcaa cgcaacttca accgttatgc cgtctttctc gccttcgata taaatcggct 780
cttcatgaac gacttctttg gaacggttta agtactcaac atagcttttg attccgcctt 840
cgtagtggta ctcgtttttc cgttcttgtc cttcacgttt gtcttcaatc gtgatgttta 900
cgccttttgt caggaaggcc aattcccgga cacggtttga aagcagatca tagtcgtata 960
cgattgtttc tttgaagatt tccggatccg gaacgaagtg cgtaatcgtt ccggtcttat 1020
cagtatcacc gatcacttca agatcagcca caggtacacc gcgctcgtac gcctgatagt 1080
ggatttttcc gtcacgatga accgtaacgt caagagtggt cgacaaggcg tttacgacgg 1140
acgcccctac accgtgaaga ccgccggata catatatccg ctgcagtatc cgc 1193
Claims (6)
- A kind of 1. bacillus amyloliquefaciens (Bacillus amyloliquefaciens) EZ10-01, on December 26th, 2016 In China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, preserving number is CGMCC No. 13495.
- A kind of 2. biocontrol agent prepared by bacillus amyloliquefaciens EZ10-01 as described in claim 1.
- 3. biocontrol agent according to claim 2, it is characterised in that be prepared by the following method to obtain:By EZ10-01 bacterium Strain is seeded on LB culture mediums, is incubated overnight, then the EZ10-01 bacterial strains of activation are transferred in Landy culture mediums and shaken Bottle fermentation, temperature are 30 DEG C, and shaking speed is 200 rpm, and fermentation time 35-40 hours to bacterium amount reach 2.46 × 109 CFU/ mL；Wherein Landy culture mediums, which form, is：20 g glucose, 5 g Pidolidones, 0.5 g MgSO4, 0.5 g KCl, 1 g KH2PO4, 0.15 mg Fe2SO4·6H2O, 5.0 mg MnSO4·H2O, 0.16 mg CuSO4·5H2O, pH7.0,1000 mL Distilled water.
- 4. the bacillus amyloliquefaciens EZ10-01 described in claim 1 is suppressing corn bacterial brown rot germ, rice bacterial leaf spot Application in germ, xanthomonas oryzae pv. oryzicola, fusarium graminearum and/or corn wheel branch Fusariumsp.
- 5. according to the application described in right 4, it is characterised in that application process is that fermented and cultured obtains zymotic fluid, and zymotic fluid is carried out Foliage-spray after dilution.
- 6. the bacillus amyloliquefaciens EZ10-01 described in claim 1 is white in preparation preventing and treating corn bacterial brown rot germ, rice Leaf spoting bacteria, xanthomonas oryzae pv. oryzicola, fusarium graminearum and/or corn wheel branch Fusariumsp biocontrol agent in should With.
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