CN105176894B - A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention graw mold of tomato - Google Patents
A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention graw mold of tomato Download PDFInfo
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Abstract
The invention belongs to Strategies of Agricultural Bio-control fields, specifically disclose a kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) bacterial strain HMB27688 for preventing graw mold of tomato, and deposit number is CGMCC No.11457.The invention also discloses using the bacterium prepare microbial bacterial agent and they prevention graw mold of tomato on application.The present invention provides an efficient microorganism for prevention graw mold of tomato, and average preventive effect is and with strong points 80.0% or more；Secondly, microbial bacterial agent of the present invention belongs to environmentally friendly to people, animal safety；In addition, it is long using the present invention prevention graw mold of tomato lasting period, it is not likely to produce drug resistance；Preparation method of the present invention is simple, it is at low cost, using simple.
Description
Technical field
The invention belongs to Strategies of Agricultural Bio-control fields, and in particular to a kind of solution starch gemma bar of prevention graw mold of tomato
Bacterium, and the microbial bacterial agent containing the bacillus amyloliquefaciens further relate to their applications on prevention graw mold of tomato.
Background technology
Graw mold of tomato is a kind of important disease caused by the pathogen of Botrytis cinerea (Botrytis cinerea Pers.).It should
Disease main harm fruit, causes fruit rot, also endangers leaf, stem and flower, up to 20%~50%, loss is tight for the tomato underproduction
Weight；Spread speed is very fast after the onset of the disease, seriously threatens tomato in greenhouse production.Its pathogen host range is wider, except invading
It contaminates outside solanaceous vegetable, can also infect the crops such as melon, wild cabbage, Kidney bean, lettuce, onion, apple.
Currently, the main control method to graw mold of tomato has：First, chemical prevention.Though chemical prevention has quick, effect
The advantages that rate is high, but because be used for a long time can caused by environmental pollution, endanger the shortcomings of natural enemy of pest, therefore be gradually eliminated；
Followed by biological control, since biological control drug effect is high, the lasting period is long, is not likely to produce drug resistance, advantages of environment protection increasingly
It has been favored by people.
Bacillus (Bacillius sp) is a kind of biocontrol bacteria received significant attention, wide, easily separated with its distribution
Culture can generate the stronger gemma of resistance, the features such as storage period is long and easy to use, become a kind of micro- life of ideal biological and ecological methods to prevent plant disease, pests, and erosion
Object.Because bacillus can generate endogenous spore, there is extremely strong anti-adversity ability, compares other kinds of bio-control factors, be more advantageous to
The production of microbial inoculum is survived in formulation environment, colonizes and is bred.Therefore, the bud inhibited to pathogen is screened
Spore bacillus is most effectual way one of of the prevention in relation to plant disease.Bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens) it is one of bacillus for preventing graw mold of tomato, is presently used for prevention graw mold of tomato
Bacillus amyloliquefaciens mainly have the CGMCC No.4777 (patent No.s：ZL201210059785.0), BA-KA3 (patent applications
Number：201410618581.5), SZ23 (number of patent application：201410683826.2), LH-1 (number of patent application：
201410735508.6) and NCPSJ17 (number of patent application：201310112580.9) etc..Due to the diversity of pathogen and same
Stepping characteristic, it is therefore desirable to find different new strains prevention graw mold of tomato.
Invention content
Present invention aims at a kind of Bacillus amyloliquefaciens strain of prevention graw mold of tomato is provided, which has height
Effect, broad spectrum antibacterial activity advantage.
The present invention second is designed to provide a kind of microbial bacterial agent produced using above-mentioned bacillus amyloliquefaciens.
Third of the present invention is designed to provide the preparation method of mentioned microorganism microbial inoculum.
The present invention the 4th is designed to provide purposes of the above-mentioned Bacillus amyloliquefaciens strain on prevention graw mold of tomato.
The present invention the 5th is designed to provide purposes of the mentioned microorganism microbial inoculum on prevention graw mold of tomato.
The present invention the 6th is designed to provide the identification method of above-mentioned Bacillus amyloliquefaciens strain.
The present invention the 7th is designed to provide the identification method of mentioned microorganism microbial inoculum.
The invention is realized by the following technical scheme：
A kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) bacterial strain HMB27688, oneself was in 2015
September is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on the 29th, and (preservation address is：Court of Beijing
The institute 3 of positive area's North Star West Road 1), deposit number is CGMCC No.11457.
The microbial bacterial agent produced using above-mentioned bacillus amyloliquefaciens HMB27688, active constituent are HMB27688 bacterium
Body.
Mentioned microorganism microbial inoculum can be liquid preparation.
Mentioned microorganism microbial inoculum, the wherein viable count of HMB27688 are more than 18.0 × 108cfu/mL。
The preparation method of mentioned microorganism microbial inoculum, includes the following steps：
(1) actication of culture：The HMB27688 bacterial strains of Cord blood are activated on LB plating mediums, picking single bacterium colony exists
It is cultivated 10~16 hours for 25~35 DEG C on LB slant mediums, obtains the bacterial strain of activation；
(2) prepared by seed liquor：The inoculation of a ring step (1) activation is scraped to 100mL LB liquid with sterile oese
In body culture medium, in 25~35 DEG C, shaking speed to be cultivated 10~16 hours under conditions of 150~220rpm, seed liquor is obtained；
(3) fermented and cultured：The seed liquor of step (2) is linked into corn flour soya bean by the ratio for being 1~3% according to volume ratio
In powder culture medium, 40~50h of fermented and cultured under conditions of temperature is 25~35 DEG C, shaking speed is 150~220rpm must be sent out
Zymotic fluid；
(4) thalline and brood cell's quantity in zymotic fluid are detected, waits in zymotic fluid that ripe brood cell accounts for brood cell and thalline sum
Stop fermented and cultured when 90%；Gained is the liquid preparation of HMB27688 bacterial strains.
LB plating mediums, LB slant mediums and the LB liquid medium is conventionally prepared.
LB plating mediums described in above-mentioned preparation method step (1) or LB slant mediums, constituent and its
Weight ratio is：8~12g of tryptone, 4~6g of yeast extract, 4~6g of sodium chloride, 12~18g of agar powder, water 1000mL.
LB liquid medium described in above-mentioned preparation method step (2), constituent and its weight ratio are：Tryptose
8~12g of peptone, 4~6g of yeast extract, 4~6g of sodium chloride, water 1000mL.
Corn flour soybean powder medium described in above-mentioned preparation method step (3), constituent and its weight percent
Than for：Corn flour 1.0~3.0%, analysis for soybean powder 1.0~3.0%, NaCl 0.1~1.0%, MnSO4·H2O 0.5~1.0%,
Remaining is water；PH value is 7.2.
The preparation method of the corn flour soybean powder medium, according to weight percent by corn flour, analysis for soybean powder, NaCl
And MnSO4·H2O is mixed, and is added water, and is adjusted pH and is stirred evenly.
Applications of the above-mentioned bacillus amyloliquefaciens HMB27688 on prevention graw mold of tomato.
Application of the mentioned microorganism microbial inoculum on prevention graw mold of tomato.
The application method of microbial bacterial agent of the present invention：Above-mentioned gained microbial bacterial agent, which is diluted with water to viable bacteria body number, is
107Cfu/mL carries out foliar spray before the onset of graw mold of tomato.
The screening separation process of HMB27688 bacterial strains
HMB27688 bacterial strains are Cotton Soil of the Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie from Jingzhou City of Hubei Province
In it is isolated.In April, 2013, Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie's Plant Protection Institute was adopted for 5 points from Jingzhou City of Hubei Province cotton field
Collect soil sample, weighing 1g after mixing is put into the sterilizing triangular flask of 250mL, and 100mL sterile waters are added, are put on shaking table, 170r/
Min vibrates 30min, stands 2h, and supernatant 10mL is taken to be added in 50mL sterile centrifugation tubes, in 80 DEG C of waters bath with thermostatic control 30 minutes, so
After take 1mL to add sterile water 9mL, 10mL10-3Times edaphon suspension, is then diluted to 10 by soil supension-4、10-5、
10-6Times dilution, takes each 200 μ L of concentration microorganism suspension to be applied on LB culture medium flat plates, each concentration is repeated 3 times, at 30 DEG C
Constant temperature incubation 1d-3d carries out the separation and purifying of bacterium.And using graw mold of tomato as target, pass through tablet face-off method, tooth in vitro
Piece method, pot experiment method carry out the screening of biocontrol microorganisms.As a result therefrom filtering out one there is apparent prevention to imitate graw mold of tomato
The bacterial strain of fruit is named as HMB27688.
The taxonomic identification of HMB27688 bacterial strains：
(1) identification by morphological characters
It is rod-shaped that thalline is cultivated on LB culture mediums, and brood cell is generated after cultivating 10h, raw in brood cell, ellipse, and cyst is not swollen
Greatly, acid-fast stain is negative, and no parasporal crystal can move, flagellum Zhousheng.On nutrient agar panel, the light breast of Initial stage of culture bacterium colony
White, purulence shape is round, neat in edge, and bacterium colony protuberance is in steamed bun shape, surface wettability；Late stage of culture bacterium colony is faint yellow, and edge is not whole
Together, dry tack free has fold；It crosses and cultivates on nutrient agar slopes, it is linear；Static gas wave refrigerator in liquid medium, table
Face forms white mycoderm.These morphological features with《Common bacteria system identification handbook》(east show pearl etc. writes Science Presses
.2001 year) described in bacillus morphological feature it is almost the same, tentatively judge that bacterial strain HMB27688 belongs to bacillus
(Bacillus)。
(2) 16S rDNA Sequence Identifications are utilized to classify
Using the genomic DNA of HMB27688 as template, PCR amplification is carried out by primer pair 16S rDNA of F27 and R1492,
The primer sequence is：
F27：5’AGAGTTTGATCATGGCTCAG3’；(SEQ ID No:1),
R1492：5’GGCTACCTTGTTACGACTT3’；(SEQ ID No:2)；
The amplification reaction system of 16S rDNA is 50 μ L:10×PCR Buffer(Mg2+)5μL；dNTP Mixture
(2.5mM)5μL；Taq (5U/ μ L) 1 μ L, F27 (10 μm of ol/L) 1 μ L, R1492 (10 μm of ol/L) 1 μ L；The genome of HMB27688
DNA 50ng；ddH2O complements to 50 μ L.The reaction condition of PCR is 95 DEG C of 5min；95 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 1.5min,
30 cycles；72℃10min.Gained pcr amplification product is subjected to gel electrophoresis, delivers Shanghai Sheng Gong bioengineering Co., Ltd
Sequencing, obtains the 16S rDNA sequences of HMB27688 (see SEQ ID No:3).The 16S rDNA sequences of gained HMB27688 are existed
Carry out tetraploid rice in Genbank, the 16S rDNA homologys of results strain HMB27688 and bacillus reach 99%；
Phylogenetic tree construction (see Fig. 1), HMB27688 are aggregated to together with bacillus, illustrate that HMB27688 belongs to bacillus
Belong to (Bacillus).
(3) the identification classification of gyrB gene orders is utilized
Using HMB27688 genomic DNAs template, bacillus gyrB gene degenerate primers gyrB-F and gyrB-R are utilized
PCR amplification is carried out for primer, obtains pcr amplification product；The sequence of the wherein described primer is：
gyrB-F：5’TTGRCGGHRGYGGHTATAAAGT3’；(SEQ ID No:4)
gyrB-R：5’TCCDCCSTCAGARTCWCCCTC3’；(SEQ ID No:5)
The pcr amplification reaction system of gyrB is 50 μ L:10×PCR Buffer(Mg2+)5μL；dNTP Mixture
(2.5mM)5μL；Taq(5U/μL)1μL；GyrB-F (10 μm of ol/L) 1 μ L, gyrB-R (10 μm of ol/L) 1 μ L；HMB27688 genes
Group DNA 50ng；ddH2O complements to 50 μ L.The reaction condition of PCR is 95 DEG C of 5min；95 DEG C of 30s, 55 DEG C of 45s, 72 DEG C of 1min,
30 cycles；72℃10min.Amplified production is delivered into the sequencing of Shanghai Sheng Gong bioengineering Co., Ltd, obtains HMB27688 bacterial strains
GyrB gene orders (see SEQ ID No:6).By the gyrB gene orders of the HMB27688 bacterial strains of acquisition in Genbank into
Row tetraploid rice, at the same using MEGA softwares (Molecular Evolutionary Genetics Analysis, molecule into
Change genetic analysis) phylogenetic tree construction.As a result, it has been found that the gyrB gene orders of HMB27688 and bacillus amyloliquefaciens are homologous
Property highest；Phylogenetic tree construction (see Fig. 2), HMB27688 bacterial strains are aggregated to together with bacillus amyloliquefaciens, explanation
HMB27688 is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and is a new strains.
In summary morphological feature, 16S rDNA and gyrB gene homology comparative analyses as a result, understanding
HMB27688 belongs to bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and with existing solution starch gemma
Bacillus strain is different, is a new strains.
The identification method of above-mentioned Bacillus amyloliquefaciens strain HMB27688 includes being with the genomic DNA of strain to be tested
Template carries out PCR amplification by primer pair of F27 and R1492, if gained pcr amplification product is SEQ ID No:Core shown in 3
Nucleotide sequence, as HMB27688 bacterial strains.
The identification method of above-mentioned Bacillus amyloliquefaciens strain HMB27688 includes being with the genomic DNA of strain to be tested
Template carries out PCR amplification by primer of gyrB-F and gyrB-R, if gained pcr amplification product is SEQ ID No:Shown in 6
Nucleotide sequence, as HMB27688 bacterial strains
The identification method of mentioned microorganism microbial inoculum includes the genomic DNA to be extracted in microbial inoculum to be measured as template, with F27
It is that primer pair carries out PCR amplification with R1492, if gained pcr amplification product is SEQ ID No:Nucleotide sequence shown in 3,
As HMB27688 microbial bacterial agents；Or the genomic DNA to be extracted in microbial inoculum to be measured is with gyrB-F and gyrB-R as template
Primer carries out PCR amplification, if gained pcr amplification product is SEQ ID No:Nucleotide sequence shown in 6, as HMB27688
Microbial bacterial agent.
The present invention has the advantage that and advantageous effect：(1) present invention provides one efficiently for prevention graw mold of tomato
Microorganism opens a new controlling way；(2) medicines of the bacillus amyloliquefaciens HMB27688 of the present invention to graw mold of tomato
Effect is high, and average preventive effect is and with strong points 80.0% or more；(3) microbial bacterial agent of the present invention belongs to environment to people, animal safety
Friendly；(4) it is not likely to produce drug resistance using the method for the present invention prevention graw mold of tomato；(5) preparation method of the present invention it is simple,
It is at low cost, using simple.
Description of the drawings
Fig. 1 are the systematic growth tree graph of the HMB27688 bacterial strains obtained according to 16S rDNA sequences.
Fig. 2 are the systematic growth tree graph of the HMB27688 bacterial strains obtained according to gyrB gene orders.
Specific implementation mode
The present invention is further clearly explained with specific embodiment below, but is not constituted in any way to the present invention
Limitation.Experimental method in following embodiments is unless otherwise instructed conventional method；Percentage in following embodiments contains
Amount, is weight percentage unless otherwise instructed.
The preparation of embodiment 1HMB27688 microbial bacterial agents
It carries out in accordance with the following steps：
(1) actication of culture：By be stored in -80 DEG C bacterial strain HMB27688 (Bacillus amyloliquefaciens strain HMB27688 oneself
It was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center in 29th in September in 2015, deposit number is
CGMCC No.11457) in LB plating mediums, (its constituent and its weight ratio are：Tryptone 10g, yeast extract
5g, sodium chloride 5g, agar powder 15g, water 1000mL) on activated at 30 DEG C, picking single bacterium colony is in LB slant mediums (its group
It is at ingredient and its weight ratio：Tryptone 10g, yeast extract 5g, sodium chloride 5g, agar powder 15g, water 1000mL) on
It is cultivated 12 hours at 30 DEG C, obtains the bacterial strain of activation.
(2) preparation of seed liquor：Making LB liquid medium according to a conventional method, (its constituent and its weight ratio are：Pancreas
Peptone 10g, yeast extract 5g, sodium chloride 5g, water 1000mL), it is packed into LB liquid medium in 250mL triangular flasks
100mL, high pressure moist heat sterilization access the bacterial strain activated in an oese step (1) after temperature drops to room temperature, in every bottle,
30 DEG C, shaken cultivation is carried out under conditions of shaking speed 180rpm 12 hours, obtain seed liquor.
(3) preparation of corn flour soybean powder medium：According to weight percent by corn flour 1.5%, analysis for soybean powder 2.0%,
NaCl 0.5%, MnSO4·H2O 0.6% is added to the water, and is uniformly mixed to get corn flour soybean powder medium；It is sub-packed in
In 500mL triangular flasks, every bottle of 200mL；Sterilizing 30 minutes is carried out to corn flour soybean powder medium at 121 DEG C, then cools to 30
It is DEG C spare.
(4) fermented and cultured：Into every bottle of corn flour soybean powder medium 200mL obtained by step (3) obtained by inoculation step (2)
Seed liquor 2mL；Fermented and cultured 36 hours is carried out under the conditions of 30 DEG C, shaking speed 180rpm, later every 30 minutes from triangle
Bottle in sampling carry out microscopy, in the visual field brood cell and total thalline number count, and calculate brood cell lead (brood cell lead (%)=
Ripe brood cell's number/(ripe brood cell's number+thalline number) × 100)；Brood cell, which leads, stops fermented and cultured when reaching 90%；Common fermentation culture
48h obtains the liquid preparation of bacillus amyloliquefaciens HMB27688.
Antagonism tests of the 2 bacillus amyloliquefaciens HMB27688 of embodiment to tomato gray mould bacterium
(1) test period and place：In early August, 2013 is in Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie plant
It is carried out in biological control of diseases laboratory.
(2) test method：
(1) for examination tomato gray mould bacterium source：Tomato gray mould bacterium BC-1 bacterial strains pick up from Hebei province Baoding Rongcheng County Dong Niudong
Village village tomato disease fruit, isolates and purifies through Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie, Agricultural University Of Hebei's Plant Protection
Department of plant pathology of institute is accredited as the pathogen of Botrytis cinerea (Botrytis cinerea), and Pathogenic Tests show as High pathogenicity.
(2) tablet face-off experiment：
Tomato gray mould bacterium BC-1 is activated on PDA plate first, in colony edge region after culture 4 days, with card punch () bacterium piece is made in punching, by the switching of tomato gray mould bacterium bacterium piece in another PDA plate center, then by 1 step of embodiment
(1) the bacillus amyloliquefaciens HMB27688 points activated in are connected on away from 2.0 centimeters of indicator bacteria bacterium piece, if blank control (not point
Connect the tomato gray mould bacterium growing state of HMB27688 bacterial strains).25 DEG C of constant temperature incubations wait for that blank control will cover with entire culture dish
When, measure control increment (colony radius) and (the inhibition growth after inoculation HMB27688 of processing increment of tomato gray mould bacterium
Radius), antagonism is indicated with bacteriostasis rate.Calculation formula is：
Bacteriostasis rate (%)=(control increment-processing increment)/control increment × 100
As a result the bacillus amyloliquefaciens HMB27688 that (is shown in Table 1) is to the inhibiting rate of tomato gray mould bacterium up to 88.16%；Transparent
11.5 millimeters of antibacterial bandwidth；Illustrate that bacillus amyloliquefaciens HMB27688 of the present invention is strong to tomato gray mould bacterium biocidal property, has anti-
Control the Biocontrol Potential of graw mold of tomato.
Antagonism test result of the table 1HMB27688 bacterial strains to tomato gray mould bacterium
| Strain name | Compare increment (mm) | Handle increment (mm) | Bacteriostasis rate (%) |
| HMB27688 | 38.0 | 4.5 | 88.16 |
Contrast tests of the 3 bacillus amyloliquefaciens HMB27688 of embodiment to graw mold of tomato control effect
(1) test process：
(1) microbial bacterial agent：HMB27688 liquid preparations prepared by embodiment 1；It is diluted with water 50 times.
(2) chemical agent：Fluoxastrobin suspending agent (250 grams per liter) (Syngenta Co., Ltd of Britain)；It is diluted with water 500 times.
(3) blank control：Clear water
(2) test method：This experiment in Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie's disease flocking biocontrol laboratory into
Row.Tomato seedling is cultivated in 50 hole seedlings nursing plates, and (tomato L-402, Liaoning gardening seedling company, Liaoning Academy of Agricultural Sciences's gardening are ground
Study carefully and produced), it is transplanted to when growing 2 compound leaves in the flowerpot of 20 centimetres of diameter, per 1 seedling of basin.Plant strain growth is to 7-8 piece compound leaves
When, select leaf age consistent, blade of the same size first uses aseptic water washing leaf table, aseptic filter paper suck dry moisture, then in embodiment
Rear (suspension mycetome 10 is diluted with water in 1 gained7Cfu/mL it is dipped in bacillus amyloliquefaciens HMB27688 microbial inoculums),
Make leaf surfaces fully medicine, be then placed in the culture dish for being covered with dual-layer sterilization filter paper, while accessing tomato gray mould bacterium BC-1
Bacterium disk (diameter 6mm), moisturizing culture.The blade handled using unused HMB27688 microbial inoculums is blank control.Wait for that blank control is abundant
Lesion diameter is investigated after the onset, calculates control effect.
Comparative test results of the 2 bacillus amyloliquefaciens HMB27688 of table to graw mold of tomato preventive effect
| Processing | Lesion diameter (mm) | Preventive effect (%) |
| HMB27688 liquid preparations | 4.0c | 83.33 |
| Chemical agent | 6.8b | 71.67 |
| Blank control | 24.0a | -- |
As a result the excised leaf that (is shown in Table 2) life survey the result shows that, bacillus amyloliquefaciens HMB27688 of the present invention is to tomato ash
Mildew preventive effect is 83.33%, and the preventive effect of chemical agent is that 71.67%, HMB27688 is apparently higher than the preventive effect of graw mold of tomato
It is good to illustrate that bacillus amyloliquefaciens HMB27688 and its microbial bacterial agent of the present invention have graw mold of tomato for chemical agent
Control effect.
Contrast tests of the 4 bacillus amyloliquefaciens HMB27688 of embodiment to graw mold of tomato control effect
(1) test process：
(1) microbial bacterial agent：HMB27688 liquid preparations prepared by embodiment 1；It is diluted with water 50 times.
(2) chemical agent：Fluoxastrobin suspending agent (250 grams per liter) (Syngenta Co., Ltd of Britain)；It is diluted with water 500 times.
(3) blank control：Clear water
(2) test method：In December, 2013 is in Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie's artificial climate room
It carries out.Tomato seedling (tomato L-402, Liaoning gardening seedling company, Liaoning Academy of Agricultural Sciences's gardening research are cultivated in heliogreenhouse
Produced), it waits for that tomato growth goes out 8 compound leaves, moves to temperature and humidity control culturing room, choose tomato seedling of the same size, by embodiment 1
50 times of water diluents of the HMB27688 liquid preparations of preparation are uniformly sprayed on tomato seedling, cultivate 24 hours at 25 DEG C, then
Access tomato gray mould bacterium bacterium disk (cultivating 3-5d, diameter 6mm at 25 DEG C on PDA plate) on each leaflet, moisturizing culture.If clear
Water and chemical agent compare, as a result, measuring each lesion diameter (A, B) using cross mensuration, calculating is sick for investigation morbidity after 3d
Spot area and control effect.
Lesion area (mm2)=π × A × B/4
Control effect (%)=(control lesion area-processing lesion area)/control lesion area × 100
As a result the live body that (is shown in Table 3) life survey the result shows that, bacillus amyloliquefaciens HMB27688 significantly suppresses tomato gray mould
The effect of extension of the bacterium on tomato leaf, prevention graw mold of tomato is 86.70%, and the preventive effect of chemical agent is
73.02%；HMB27688 of the present invention is significantly higher than the preventive effect of graw mold of tomato the preventive effect of chemical agent.Illustrate Xie Dian of the present invention
Afnyloliquefaciens HMB27688 and its microbial bacterial agent have good control effect to graw mold of tomato.
Comparative test results of the 3 bacillus amyloliquefaciens HMB27688 of table to graw mold of tomato preventive effect
| Processing | Lesion area (mm2) | Preventive effect (%) |
| HMB27688 liquid preparations | 15.02c | 86.70 |
| Chemical agent | 30.42b | 73.02 |
| Blank control | 112.76a | -- |
Contrast tests of the 5 bacillus amyloliquefaciens HMB27688 of embodiment to graw mold of tomato control effect
(1) test process：
(1) microbial bacterial agent：HMB27688 liquid preparations prepared by embodiment 1, are diluted with water 50 times.
(2) chemical agent：Fluoxastrobin suspending agent (250 grams per liter) (Syngenta Co., Ltd of Britain)；It is diluted with water 500 times.
(3) blank control：Clear water
(2) test method：2 months-June in 2014 Hebei province Mancheng County of Baoding contest village Sun Mao dragon tomato greenhouses into
Row.Start tomato leaf portion gray mold moderate generation in entire canopy when experiment, sick leaf is extractd before dispenser.Per three row of cell, four weights
It is multiple, random district's groups arrangement.Using the bacillus amyloliquefaciens HMB27688 prepared by knapsack electric sprayer inoculation embodiment 1
Microbial inoculum (is diluted, mycetome 10 with water7cfu/mL)；Separately set comparison medicament and blank control.Dispenser for the first time is sprayed again after 7 days
It applies 1 time, the single plant disease number of sheets of each processing cell of the 7th after secondary dispenser day investigation, and calculates preventive effect.
As a result after the bacillus amyloliquefaciens HMB27688 microbial inoculums that (are shown in Table 4) processing, the single plant disease number of sheets of graw mold of tomato
Substantially less than blank control and chemical agent control, preventive effect reaches 85.72%, illustrates HMB27688 pairs of bacillus amyloliquefaciens
Graw mold of tomato has good control effect.
Comparative test results of the 4 bacillus amyloliquefaciens HMB27688 of table to graw mold of tomato preventive effect
| Processing | The single plant disease number of sheets (piece) | Preventive effect (%) |
| HMB27688 liquid preparations | 0.67c | 85.72 |
| Chemical agent | 1.33b | 71.52 |
| Blank control | 4.67a | --- |
Claims (5)
1. a kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) bacterial strain HMB27688, oneself was in 2015 9
The moon is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number CGMCC on the 29th
No.11457。
2. utilizing the microbial bacterial agent of bacillus amyloliquefaciens HMB27688 productions described in claim 1, it is characterised in that its
Active constituent is HMB27688 thalline.
3. microbial bacterial agent according to claim 2, it is characterised in that the microbial bacterial agent is liquid preparation.
4. applications of the bacillus amyloliquefaciens HMB27688 described in claim 1 on prevention graw mold of tomato.
5. application of the microbial bacterial agent according to claim 2 or 3 on prevention graw mold of tomato.
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