CN109306336A - Using colony induction signaling molecule AHLs as the disease control bacterial strain of target and its application - Google Patents
Using colony induction signaling molecule AHLs as the disease control bacterial strain of target and its application Download PDFInfo
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Abstract
The invention discloses a kind of using colony induction signaling molecule AHLs as the disease control bacterial strain of target, and the bacterial strain deposit number is GDMCC 60434.The bacillus cereus has preferable degrading activity for colony induction signaling molecule AHLs, and degradation effect is stable and significant, there is huge application potential in terms of prevention and treatment AHLs mediates pathogenic pathogenic bacteria harm, this provides a kind of new approaches and methods for the biological control of disease.
Description
Technical field
The present invention relates to a kind of prevention and treatment bacterial strains more particularly to a kind of using colony induction signaling molecule AHLs as the disease of target
Prevent and treat bacterial strain and its application.
Background technique
In agricultural production, efficiently prevent and treat pathogen, reduce loss caused by disease, be always the producer think it is to be achieved
Target.Pectinatus (Pectobacterium) is that great a kind of pathogen is endangered in agricultural production, and this kind of pathogen can be with
A variety of hydrolases are secreted, such as: pectase, poly galactase, protease etc..Pectinatus is posted using these hydrolases to break
The defense system of main plant, destruction organization structure of the plant, the related symptoms for successfully infecting and host plant being caused soft rot occur,
Eventually lead to the related crop species underproduction, or even total crop failure.For this kind of pathogenic bacteria, highly effective control there is no to arrange in production
It applies.
Currently, mainly passing through sprinkling chemical reagent for control measure used in Pectinatus both at home and abroad, such as: thiophene bacterium
The chemical pesticides such as ketone, thiophene, agricultural streptomycin or Zhongshengmycin or antibiotic are prevented and treated, however these chemicals
A large amount of use have had resulted in environmental pollution, and the ecological balance is destroyed, pathogen drug resistance enhances and food safety etc. is a series of
Problem.Therefore, it is extremely urgent to find novel, green, environmentally friendly, efficient pest control method.
Bacterium can synthesize, and secrete and perceive signal specific molecule, after the population density of bacterium reaches certain threshold value, ring
Signaling molecule concentration in border also reaches certain level, and at this moment certain genes of bacterium start to express, what some population levels reached
Behavior starts to occur, this phenomenon, that is, quorum sensing (Quorum sensing, QS).(Fuqua WC,SC Winans,EP
Greenberg et al.Quorum sensing in bacteria:the LuxR-LuxI family of cell
density-responsive transcriptional regulators.[J]J Bacteriol,1994,176:269).Group
Body induction is widely present in Pectinatus (Pectobacterium), Pseudomonas aeruginosa (Pseudomonas aeruginosa)
In the Gram-negative bacterias such as rice base corruption bacterium (Dickeya zeae), and participate in some important biological functions, such as water
Solve synthesis, the pyocyanin etc. of enzyme.Acyl homoserine lactones substance (N-Acylhomoserine lactones, AHLs)
For the colony induction signaling molecule of Gram-negative bacteria, AHLs is in addition to including some traditional AHLs, such as: N- (3- oxo oneself
Acyl)-L- homoserine lactone (N- (3-oxohexanoyl)-L-homoserine lactone, OHHL), (3- oxo is pungent by N-
Acyl)-L- homoserine lactone (N- (3-oxooctanoyl)-L-homoserine lactone, OOHL) etc., further include
Novel AHLs, such as: isovaleryl-homoserine lactone (Isovaleryl-homoserine lactone), carboxylation acyl group-
Homoserine lactone (carboxyl-AHLs), aryl-homoserine lactone (Aryl-homoserine lactone) and tonka-bean
Acyl-homoserine lactone (p-coumaroyl-HSL) etc..It is a kind of that (Quorum quenching, QQ), which is quenched, in quorum sensing
The mode of intervention school-based is interfered, it is by inhibiting the synthesis of signaling molecule, accumulation, monitoring or carrying out to signaling molecule
Enzyme degradation or modification mechanism come realize interference intervention school-based purpose (Fetzner S.Quorum quenching
enzymes[J].J Biotechnol,2015,201:2-14.)。
It is a kind of disease control new strategy that quorum sensing, which is quenched, and the prevention and treatment of pathogen, Bu Huizeng are carried out in the way of this
The survival pressure for adding pathogenic bacteria avoids the appearance of its drug resistance.This new way is with easy to operate, economical and practical, environment is friendly
The advantages that good.The research and development that preparation is quenched in unlike signal quorum sensing are current international research hot spots.
Summary of the invention
The present invention overcomes the deficiencies of existing pathogen biological prevention, and providing one kind can be with efficient degradation quorum sensing
The bacillus cereus (Bacillus cereus) of signaling molecule AHLs and its application in terms of disease control.Waxy gemma
Bacillus has significant degradation to colony induction signaling molecule AHLs, has in terms of prevention and treatment relies on the pathogenic pathogen of AHLs
There is great potential, to solving the problems, such as that chemical pesticide or abuse of antibiotics and pathogenic bacteria resistance to drugs have Great significance.
In order to achieve the above objectives, a kind of technical solution that the present invention uses are as follows:
It is a kind of using colony induction signaling molecule AHLs as the disease control bacterial strain of target, which is characterized in that the bacterial strain is protected
Hiding number is GDMCC 60434.
It is a kind of using colony induction signaling molecule AHLs be the disease control bacterial strain of target degradation colony induction signaling molecule
Application in AHLs, or the application in preparation degradation AHLs product.
In a preferred embodiment of the present invention, further comprise, the colony induction signaling molecule AHLs include with
It is one or more of lower: N- (3- oxohexanoyl)-L- homoserine lactone, N- (3- oxo decoyl)-L- homoserine lactone,
Isovaleryl-homoserine lactone, carboxylation acyl-homoserine lactone, aryl-homoserine lactone, coumaric acyl-Kosé ammonia
Acid lactone.
A kind of bacillus relies on AHLs in prevention and treatment and mediates application in pathogenic plant disease, or relies on AHLs in preparation and cause a disease
Pathogenic bacteria prevention and treatment preparation in application.
It in a preferred embodiment of the present invention, further comprise that the bacterial strain is Bacillus cereus strain XN-
42。
A method of prevention and treatment relies on the pathogenic pathogenic fungus diseases of AHLs, with Bacillus cereus strain XN-42 to crop
Carry out inoculation processing.
It in a preferred embodiment of the present invention, further comprise that the dependence AHLs pathogenic pathogenic bacteria include: enlightening
Cattell bacterium, Pectinatus, Pseudomonas aeruginosa.
It in a preferred embodiment of the present invention, further comprise that the degradation bacterial agent includes bacillus cereus bacterium
Strain XN-42, Bacillus cereus strain XN-42 bacterium solution.
A kind of biocontrol agent relying on the pathogenic pathogenic bacteria of AHLs, the biocontrol agent include bacillus cereus bacterium
Strain XN-42, Bacillus cereus strain XN-42 bacterium solution.
Bacterial strain of the invention is separated from the soil for picking up from Guangzhou Agricultural University Of South China school farm vegetable field through artificial screening
Purifying obtains, morphological feature and 16S rDNA Phylogenetic Analysis by the bacterial strain, identifies that strain X N-42 is waxy bud
Spore bacillus (Bacillus cereus), and it is named as XN-42.
It is preserved in Guangdong Province's Culture Collection, deposit number is GDMCC NO:60434.
The colonial morphology feature of strain X N-42: light yellow, round, protrusion, opaque, neat in edge.
The morphological feature of Electronic Speculum observation thallus are as follows: cell is in the shape of a rod, amphitrichous.
Strain X N-42 to the resistance of ampicillin, kanamycins, neomycinsulphate and streptomysin reach 400 μ g/mL with
On, 150 μ g/mL are reached to the resistance of gentamicin, 350 μ g/mL are reached to the resistance of carbenicillin, tetracyclin resistance is reached
To 10 μ g/mL, to the resistance of chloramphenicol up to 5 μ g/mL or less.
Bacillus cereus (Bacillus cereus) strain X N-42 provided by the invention can effectively degrade group sense
Induction signal molecule OHHL, can in the culture medium using OHHL as sole carbon source that concentration is 0.5mM normal growth, and in 96h
73% is reached to the colony induction signaling molecule degradation rate that initial concentration is 0.5mM, therefore, which mediates in prevention and treatment AHLs and cause
There is huge application potential in terms of the pathogenic bacteria of disease.
Inoculation processing is carried out to crop with the bacterium solution of Bacillus cereus strain XN-42, to prevent to rely on what AHLs caused a disease
Pathogenic bacteria are infected.
The optimum medium for preparing strain X N-42 bacterium solution is Luria-Bertani culture medium, formula are as follows: tryptone
10.0g/L, yeast extract 5.0g/L, 6.8~7.2,121 DEG C of sterilizing 20min of sodium chloride 10.0g/L, pH.
The invention solves the defect existing in the background technology, the present invention have it is following the utility model has the advantages that
The research of the invention finds that the bacillus cereus for colony induction signaling molecule AHLs there is preferable degradation to live
Property, and degradation effect is stable and significant, has huge application potential in terms of prevention and treatment AHLs mediates pathogenic pathogenic bacteria harm,
This provides a kind of new approaches and methods for the biological control of disease.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
Fig. 1 is the colonial morphology figure of strain X N-42 of the invention on LB culture medium.
Fig. 2 is the scanning electron microscope (SEM) photograph of strain X N-42 of the invention.
Fig. 3 is the Phylogenetic analysis figure of strain X N-42 of the invention.
Fig. 4 is the growing state figure of strain X N-42 of the invention in different antibiotic.
Fig. 5 is the degrading activity measurement chart of strain X N-42 of the invention to OHHL.(CK is to be not added with the blank that bacterium is quenched
Control).
Fig. 6 A is the comparative diagram of non-inoculating strain XN-42.
Fig. 6 B is high-efficient liquid phase chromatogram of the strain X N-42 to OHHL degradation 0d.
Fig. 6 C is high-efficient liquid phase chromatogram of the strain X N-42 to OHHL degradation 1d.
Fig. 6 D is high-efficient liquid phase chromatogram of the strain X N-42 to OHHL degradation 2d.
Fig. 6 E is high-efficient liquid phase chromatogram of the strain X N-42 to OHHL degradation 3d.
Fig. 6 F is high-efficient liquid phase chromatogram of the strain X N-42 to OHHL degradation 4d.
Fig. 7 is strain X N-42 of the invention using OHHL as the growth curve of sole carbon source and degradation curve figure.
Fig. 8 be strain X N-42, E.coli, B23 of the invention respectively with Z3-3 co-inoculation in potato tubers for 24 hours after
Incidence.
Fig. 9 be strain X N-42, E.coli, B23 of the invention respectively with Z3-3 co-inoculation in castock for 24 hours after hair
State of an illness condition.
Figure 10 be strain X N-42, E.coli, B23 of the invention respectively with Z3-3 co-inoculation in carrot for 24 hours after hair
State of an illness condition.
Specific embodiment
Presently in connection with drawings and examples, the present invention is described in further detail, these attached drawings are simplified signal
Figure, the basic structure of the invention will be illustrated schematically only, therefore it only shows the composition relevant to the invention.
Unless stated otherwise, agents useful for same and material are commercially available in the embodiment of the present invention.
Used culture medium and reagent are as follows in following embodiment:
Luria-Bertani culture medium (LB): tryptone 10.0g/L, yeast extract 5.0g/L, sodium chloride 10.0g/
6.8~7.2,121 DEG C of sterilizing 20min of L, pH.LB solid culture based formulas is that 1.5% (w/v) is added in liquid medium
Agar.
Basal salt media (MSM): (NH4)2SO4, 2.0g/L；CaCl2·2H2O, 0.01g/L；Na2HPO4·12H2O,
1.5g/L；KH2PO4, 1.5g/L；MgSO4·7H2O, 0.2g/L；FeSO4·7H2O, 0.001g/L；pH 6.5.
Basal medium (MM): K2HPO4, 10.5g/L；KH2PO4, 4.5g/L；(NH4)2SO4, 2.0g/L；Mannitol,
2.0g/L；Glycerol, 2.0g/L；MgSO4·7H2O, 0.2g/L；CaCl2, 0.01g/L；FeSO4, 0.005g/L；MnCl2,
0.002g/L；pH 6.5.
OHHL buying is purchased neat from Guangzhou from reagent needed for the virtuous Chemical Industry Science Co., Ltd in Shanghai, X-gal, culture medium
The companies such as Hunan, Hua Qisheng.
Separation, the screening of 1 bacillus cereus XN-42 of embodiment
1.1, soil sample acquires: acquiring the soil sample from Agricultural University Of South China school farm as microbial source.
Soil sample was acquired from Guangzhou, Guangdong Agricultural University Of South China school farm vegetable field, at this point on July 30th, 2017
Keep forging ahead sample, pack, as microbial source take back laboratory carry out bacterial strain separation.
1.2, the enrichment culture of bacterial strain: preparing basal salt media (MSM), and the MSM of 50mL is packed into 250mL triangular flask
Culture medium, 121 DEG C of sterilizing 20min, OHHL mother liquor is aseptically added after cooling, and (concentration 100mm/L, solvent are second
Nitrile), make 5 μm of ol/L of OHHL ultimate density in culture medium, 5g soil sample is then added, is placed in 30 DEG C, 200rpm shaking table culture
After 7d, it is transferred in the MSM culture medium that OHHL concentration is 10 μm of ol/L by 10% inoculum concentration sampling.The same terms culture 7d
Afterwards, then by 10% inoculum concentration sampling it is transferred in the MSM culture medium that OHHL concentration is 20 μm of ol/L, then with the same terms culture
7d.And so on, 7d a cycle constantly makes the concentration of OHHL increase to 80 μm of ol/L.
1.3, the separation of bacterial strain and pure strain isolation and purifying: is carried out using dilution, spread plate and plate streak
Change.
1mL end MSM culture solution is taken, being diluted to concentration with sterile water is respectively 10-1、10-2、10-3、10-4、10-5、10-6、10-7、10-8Culture solution, then draw the 100 μ L of culture solution of each concentration respectively, on LB solid plate be coated with uniformly,
It dries, 30 DEG C of cultures are for 24 hours.The single colonie of the different shape grown on picking plate, LB solid plate repeatedly cross culture it is pure
Change, until isolating single strain.Bacterium method is protected by bacterial strain preservation with glycerol, detects degrading activity to further experiment.
1.4, bacterial strain screening: have using reporting bacterial strain CF11 (Agrobacterium tumefaciensCF11) screening
The bacterial strain of degrading activity.
Bacterial strain to be screened is taken out from -80 DEG C of refrigerators, lines and is activated in LB solid medium tablets, at 30 DEG C
Incubator culture is for 24 hours.Picking single colonie, is seeded to LB liquid medium, at 30 DEG C, is incubated overnight, obtains under conditions of 200rpm
To bacterium solution.Take 1 OD600The thallus of value is transferred in the 1mL MSM culture medium that OHHL concentration is 20 μm of ol/L, and is uniformly mixed,
It obtains 1mL and waits for culture solution.It will be transferred to culture solution in 2mL centrifuge tube, with 30 DEG C, the CMC model of 200rpm is for 24 hours.After for 24 hours,
It takes 5 μ L reaction mixture point samples to dry to the MM agar strip top of 1cm wide, is reported later at one row of lower section point (about 13-18 point)
Accuse the bacterium solution of bacterial strain.Wherein, reporting bacterial strain is with 28 DEG C, and the condition of 200rpm is incubated overnight to muddiness, and agar strip is by X-gal concentration
Gained is cut for the MM plate of 40 μ g/mL.The agar strip for having put sample and reporting bacterial strain is placed in 28 DEG C of incubators, and is kept away
Light processing, observation experiment result after placing for 24 hours.
Interpretation of result, OHHL has diffusivity, and diffusion length is directly proportional to its concentration.When reporting bacterial strain CF11 is perceived
In environment OHHL there are when, the gene of beta galactosidase starts to express, and beta galactosidase is discharged into environment.β-
Galactosidase can be by leuco-compounds X-gal (the chloro- 3- indoles-β-D- galactoside of the bromo- 4- of 5-) enzyme in MM agar strip
Solution is that galactolipin and the dark blue bromo- 4- of color substance 5- are indigo, and the indigo bacterium colony for making entire reporting bacterial strain of the bromo- 4- of 5- becomes blue.
That is, when the OHHL contained in reaction mixture is more, the distance for becoming indigo plant from top is longer, conversely, then distance is longer.Root
The bacterial strain best to OHHL degradation effect is filtered out according to experimental result, is named as XN-42.
The identification of 2 Bacillus cereus strain XN-42 of embodiment
The present embodiment has carried out morphological feature, 16S rDNA Phylogenetic Analysis for degradation bacteria XN-42, identifies the bacterium
Strain is bacillus cereus (Bacillus cereus).It is specific as follows:
2.1 colony morphology characteristics: cultivating 48h on LB solid plate, and bacterium colony is light yellow, round, protrusion, opaque, side
Edge is neat, as shown in Figure 1；It is muddy in diffusivity in LB liquid medium, aerobic, well-grown at 30 DEG C.
2.2 morphological features: as shown in Fig. 2, thallus is in the shape of a rod, table amphitrichous.
2.3 16S rDNA sequences and phylogenetic analysis: the 16S rDNA gene order length of strain X N-42 is
1436bp has found after being compared with ncbi database (http://www.ncbi.nlm.nih.gov/), strain X N-42 with
Bacillus cereus homology with higher (> 99%), systematic evolution tree is as shown in Figure 3.
The antibiotics sensitivity of 3 strain X N-42 of embodiment is analyzed
In order to preferably study strain X N-42, the present embodiment has carried out Multiple Classes of Antibiotics sensibility to strain X N-42
Detection, concrete outcome are as shown in Figure 4.
The results showed that strain X N-42 resists ampicillin, kanamycins, neomycinsulphate and streptomysin
Property reaches 400 μ g/mL or more, reaches 150 μ g/mL to the resistance of gentamicin, reaches 350 μ g/ to the resistance of carbenicillin
ML reaches 10 μ g/mL to tetracyclin resistance, to the resistance of chloramphenicol up to 5 μ g/mL or less.This result is follow-up study choosing
Take suitable antibiotic as reference.
4 strain X N-42 of embodiment measures the degrading activity of OHHL
The present embodiment detects N-42 pairs of strain X using reporting bacterial strain CF11 (Agrobacterium tumefaciens)
The degradation effect of OHHL.
With LB solid medium tablets activated strains XN-42, plate is placed in 30 DEG C of incubators, culture is for 24 hours.Picking single bacterium
It falls and is seeded in LB liquid medium, with 30 DEG C, the condition of 200rpm is incubated overnight to obtain bacterium solution.Take 1 OD600The thallus of value
After evenly mixing with the 1mL MSM culture medium using OHHL as sole carbon source, it is transferred in 2mL centrifuge tube, obtains to culture solution,
Make 10 μm of ol/L of concentration of OHHL in MSM culture medium.With 30 DEG C, the CMC model of 200rpm is for 24 hours.After for 24 hours, 5 μ L is taken to react
Object point is mixed to agar strip top, then successively puts reporting bacterial strain bacterium solution in lower section, reporting bacterial strain is according to side described in embodiment 1
Method culture.Later, the agar strip for having put reaction mixture and reporting bacterial strain is placed in 28 DEG C of incubators, carries out being protected from light place
Reason, observation experiment result after culture for 24 hours.Wherein, agar strip is that the MM plate containing 40 μ g/mL X-gal cuts gained.CK is not
Blank control containing XN-42.
Experimental result illustrates to contain OHHL in sample as shown in figure 5, the agar strip about 1/2 of CK experimental group becomes blue；XN-
The unchanged indigo plant of the agar strip of 42 experimental groups illustrates that without OHHL in sample, i.e. OHHL is degradable by XN-42.The result shows that:
Strain X N-42 has the activity of degradation OHHL.
Embodiment 5 detects the activity of strain X N-42 degradation OHHL using HPLC
5.1, the strain X N-42 LB solid panel activation in -80 DEG C will be frozen, after 30 DEG C of cultures for 24 hours on picking plate
Single colonie is inoculated in LB liquid medium, and with 30 DEG C, the condition of 200rpm is incubated overnight to obtain bacterium solution.Take 1 OD600Value
Thallus, thallus are resuspended with the MSM culture medium of 1mL, re-suspension liquid are added in 19mL MSM basal medium, and add OHHL mother liquor
(concentration 100mmol/L) makes its ultimate density 0.2mmol/L.At 30 DEG C, cultivated under conditions of 200rpm, in 0d, 1d,
Five point in time sampling of 2d, 3d and 4d, and OHHL remaining in sample is extracted, it is indicated using the residual quantity of HPLC measurement OHHL
Degradation situation of the strain X N-42 to OHHL.
5.2, the extracting method of OHHL
It takes 7mL sample into 15mL centrifuge tube, after being centrifuged 10min with the speed of 4000rpm, takes 5mL supernatant to 50mL
In separatory funnel, and isometric ethyl acetate is added into separatory funnel, after acutely shaking 3min, stands, layering, by lower layer
Solution is transferred in graduated glass test tube, and the funnel that upper layer of extraction liquid liquid is covered with filter paper is filled into 50mL round-bottomed flask.Under
Layer solution is extracted again according to the above method.After filtrate rotary evaporated to dryness is dry, round-bottomed flask is washed in two times with chromatography acetonitrile,
It is settled to 2mL.Most afterwards through 0.45 μm of organic membrane filtration to sample injection bottle, HPLC to be used measures OHHL residual volume.
5.3, the HPLC testing conditions of OHHL
HPLC instrument model: Waters 2695.Chromatographic column: C18Reverse-phase chromatographic column (250 spectrums column (2695 method to be used).Stream
Speed is 0.5mL/min.Column temperature is 30 DEG C.Mobile phase: acetonitrile: water=70；30 (v:v).Detection wavelength is 210nm.Sample volume is
20μL。
5.4, degradation rate calculation method
Degradation rate (%)=(1-A1/A0) × 100, A1For OHHL residual concentration after degradation bacteria processing, A0After control treatment
OHHL residual concentration.
5.5, analysis of experimental results
HPLC testing result is as shown in fig. 6, the comparative diagram that figure A is non-inoculating strain XN-42, figure B, C, D, E, F are respectively
For strain X N-42 in 0d, 1d, 2d, 3d and 4d to the degradation figure of OHHL, degradation rate respectively reaches 0%, 30%, 50%,
64%, 73%, the OD of corresponding time strain X N-42600Value is respectively 0.002,0.547,0.640,0.784 and 0.683.Experiment
Show under the conditions of OHHL is with existing for sole carbon source, bacterial strain can degrade OHHL and using its growth.
As shown in Figure 7, the degradation of OHHL is positively correlated with strain growth, and in the presence of OHHL, strain growth is not stagnant
Stay the phase, quickly enter growth logarithmic phase, be the logarithmic phase of strain growth in 1d, at this time the bacterial strain to the degradation rate of OHHL most
Fastly.
6 strain X N-42 of embodiment studies the biocontrol effect of soft rot
The present embodiment is with plant soft rot disease pathogenic bacteria carrot soft rot Pectinatus carrot subspecies (Pectobacterium
Carotovorum subsp.carotovora, Pcc) for Z3-3, research strain X N-42 is to relying on the pathogenic pathogenic bacteria of AHLs
Biocontrol effect.
With LB solid medium tablets activated strains XN-42, Z3-3, E.coli and B23,30 DEG C of cultures.It chooses respectively afterwards for 24 hours
The single colonie made even on plate, is seeded in LB liquid medium, and with 30 DEG C, the condition of 200rpm is incubated overnight to bacterium solution OD600>
2.0.Wherein, bacillus thuringiensis subsp israelensis (Bacillus thuringiensis subsp.Israelensis)
B23 is known bacterial strain (Dong Y, Xu J, Li X, et al.AiiA, an enzyme that OHHL with degradation effect
inactivates the acylhom oserinelactone quorum-sensing signal and attenuates
the virulence of Erwinia carotovora[J].Proc Natl Acad Sci USA,2000,97(7):
3526-3531.), E.coli is the known bacterial strain for not having degradation OHHL.In biological and ecological methods to prevent plant disease, pests, and erosion experiment, four experimental groups will be set:
Z3-3+LB, Z3-3+E.coli, Z3-3+B23 and Z3-3+XN-42.Wherein, experimental group Z3-3+LB is blank control group；Z3-3+
E.coli is negative control group, and Z3-3+B23 is positive controls.
Fresh potato, Chinese cabbage and carrot are chosen as experimental material, material is cleaned with distilled water, wherein Ma Ling
Potato and carrot are cut into the slice of thickness about 0.3cm, and castock is cut to the small rectangle of about 6cm × 4cm.Z3-3 bacterium solution is distinguished
It is mixed in a certain proportion with the bacterium solution and LB liquid medium of B23, E.coli, XN-42, makes XN-42, E.coli, B23, Z3-3
Final work OD600=2.0.The mixed bacteria liquid of each experimental group takes 2 μ L to be seeded to the experimental material center handled well respectively.Add
Enter wet white Hupeh anemone root and leaf moisturizing and prevent living contaminants with preservative film sealing, places 28 DEG C of incubator cultures and observe result afterwards for 24 hours.
As a result as shown in Fig. 8, Fig. 9 and Figure 10, in the biological and ecological methods to prevent plant disease, pests, and erosion experiment of potato, Chinese cabbage and carrot, experimental group Z3-3+
The onset area of LB and Z3-3+E.coli is larger compared with experimental group Z3-3+B23 and Z3-3+XN-42 onset area, occurring degree compared with
It is serious.That is OHHL degradation bacteria XN-42 and pathogenic bacteria co-inoculation are obvious compared with soft rot Disease symptoms when being individually inoculated with pathogenic bacteria
Mitigate.The experimental results showed that strain X N-42 is to carrot soft rot Pectinatus carrot subspecies (Pectobacterium
Carotovorum subsp.carotovora, Pcc) bacterial soft rot of potato, soft rot of cabbage and carrot soft rot caused by Z3-3
Disease has significant biocontrol effect.
Through the foregoing embodiment the study found that the bacillus cereus has preferably for colony induction signaling molecule AHLs
Degrading activity, and degradation effect is stable and significant, with huge in terms of prevention and treatment AHLs mediates pathogenic pathogenic bacteria harm
Application potential, this provides a kind of new approaches and methods for the biological control of disease.
Based on the above description of the preferred embodiments of the present invention, through the above description, related personnel completely can be with
Without departing from the scope of the technological thought of the present invention', various changes and amendments are carried out.The technical scope of this invention
It is not limited to the contents of the specification, it is necessary to determine the technical scope according to the scope of the claims.
Claims (9)
1. a kind of using colony induction signaling molecule AHLs as the disease control bacterial strain of target, which is characterized in that the bacterial strain preservation
Number be GDMCC 60434.
2. it is a kind of using colony induction signaling molecule AHLs be the disease control bacterial strain of target degradation colony induction signaling molecule
Application in AHLs, or the application in preparation degradation AHLs product.
3. special using colony induction signaling molecule AHLs as the disease control bacterial strain of target according to claim 1 or described in 2
Sign is that the colony induction signaling molecule AHLs includes following one or more: in N- (3- oxohexanoyl)-L- homoserine
Ester, N- (3- oxo decoyl)-L- homoserine lactone, isovaleryl-homoserine lactone, carboxylation acyl-homoserine lactone,
Aryl-homoserine lactone, coumaric acyl-homoserine lactone.
4. a kind of bacillus relies on the application in the pathogenic plant disease of AHLs mediation in prevention and treatment, or relies on what AHLs caused a disease in preparation
Application in the prevention and treatment preparation of pathogenic bacteria.
5. the application according to claim 2 or 4, which is characterized in that the bacterial strain is Bacillus cereus strain XN-
42。
6. a kind of method that prevention and treatment relies on the pathogenic pathogenic fungus diseases of AHLs, which is characterized in that use Bacillus cereus strain XN-
42 pairs of crops carry out inoculation processing.
7. the method that prevention and treatment according to claim 6 relies on the pathogenic pathogenic fungus diseases of AHLs, which is characterized in that it is described according to
The pathogenic bacteria for relying AHLs pathogenic include: enlightening Cattell bacterium, Pectinatus, Pseudomonas aeruginosa.
8. a kind of degradation bacterial agent of degradable colony induction signaling molecule AHLs, which is characterized in that the degradation bacterial agent includes wax
Sample Bacillus strain XN-42, Bacillus cereus strain XN-42 bacterium solution.
9. a kind of biocontrol agent for relying on the pathogenic pathogenic bacteria of AHLs, which is characterized in that the biocontrol agent includes waxy
Bacillus strain XN-42, Bacillus cereus strain XN-42 bacterium solution.
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| CN111154703A (en) * | 2020-03-25 | 2020-05-15 | 福建农林大学 | Bacillus cereus containing quorum sensing quenching gene aiiA and application thereof |
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