CN109868250A - One plant inhibits virus, the Bei Laisi bacillus for promoting plant growth and its application - Google Patents
One plant inhibits virus, the Bei Laisi bacillus for promoting plant growth and its application Download PDFInfo
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Abstract
Of the invention one plant inhibits virus, the Bei Laisi bacillus for promoting plant growth and its application, it is characterized in that, the strain name is Bei Laisi bacillus (Bacillus velezensis) SNB55 and the bacterial strain in the prevention and treatment viroses of plant and fungal diseases of plants, promotes the application in plant growth.The beneficial effects of the present invention are: one plant of the invention inhibits virus, the Bei Laisi bacillus of plant growth and its application is promoted to have nontoxic, no pathogenicity, and to safety of human and livestock, environmentally friendly, resistance is strong, bacteriostatic activity is stable；It is able to suppress Bacillus cercus, inhibits tobacco mosaic virus (TMV) and tomato yellow leaf curl virus, inhibit the phytopathogens such as cucumber fusarium axysporum, Tobacco target spot bacterium, phytophthora blight of pepper, Rhizoctonia solani Kuhn and botrytis cinerea, heteroauxin can be secreted, promote seed sprouting and plant growth, it is one plant of multi-functional biocontrol microorganisms, has broad application prospects.
Description
Technical field
The invention belongs to microbial control technical fields, are related to a kind of microbial control technology of tomato virus disease, specifically
More particularly to one plant of inhibition tobacco mosaic virus (TMV) (Tobaco mosaic virus, TMV) and tomato yellow leaf curl virus
The Bei Laisi bacillus (Bacillus velezensis) of (Tomato yellow leaf curl virus, TYLCV) and
It is applied.
Background technique
The viroses of plant are known as the title of " plant cancer ", are the second largest plant diseases after fungal diseases of plants, give agriculture
Industry production causes heavy economic losses.Plant virus is many kinds of, wherein tobacco mosaic virus (TMV) (Tomato mosaic
Virus, TMV) it is to cause one of main malicious source of the plants of Solanaceae such as tomato and tobacco.In recent years, tomato yellow leaf curl virus
(Tomato yellow leaf curl virus, TYLCV) is propagated rapidly in China, and in the trend increased year by year, to China
Tomato production constitutes a serious threat.For a long time, the viroses of plant are prevented and treated based on chemical prevention and the antiviral kind of cultivation,
In terms of Agro-chemicals control, satisfactory anti-plant virus agent functionization kind is not much at present, and especially special efficacy is therapeutic
Medicament is less, and the medicament reported is undesirable in field practical application preventive effect, and chemical prevention is there is also many problems,
The problems such as largely applying chemical pesticides, causing pesticide residue, food safety, ecological environmental pollution.So cultivating antiviral kind
It is considered as the economic and effective approach for preventing and treating tobacco mosaic virus disease.But it is limited commercially valuable resistant material
The progress of breeding work is greatly limited, there is no the kind of high-quality highly resistance that can be widely applied at present.With the viroses of plant
Harm aggravate, the problems such as long-term a large amount of uses of chemical pesticide, environmental pollution, pesticide residue, develop new, efficient, safely
Microorganism formulation control the viroses of plant, have become the needs of agricultural safety and sustainable development.
Active strong, safe, the high-efficient advantage of antiviral active substance that microbial metabolism generates is provided from microorganism
It is screened in source, separation obtains the antiviral substances such as polysaccharide, nucleic acid, protein and small molecule and has become the hot spot studied now.Have
Formation product, play a significant role in agricultural production, wherein lentinan China be successfully registered as it is disease-resistant
Toxic agent, industrialized production.The development of especially anti-virus type farm antibiotics has achieved breakthrough, Ningnanmycin and
Cytosintetidemycin be research it is most successful be anti-virus type farm antibiotics.Ningnanmycin is isolated from the change of streptomyces noursei Xichang
Kind, it is a kind of novel cytosine nucleoside antibiotic, there is wide spectrum and efficient inhibiting effect, and low toxicity, low-residual etc. are excellent
Point, is widely used to agricultural production, and the concentration that the preventive effect to TMV infection is 500 μ g/mL is 58.1%.It is reported that Ningnan
Mycin can induce plant and increase anti-TMV activity.The cytosintetidemycin isolated from streptomyces hygroscopicus shows effective anti-plant
Virus activity, and in China, successfully registration and industry turn to anti-plant virus agent.And other most of anti-phytoviral activities
Substance and biocontrol bacterial strain are in the laboratory research stage, therefore, still lack in production for preventing and treating the novel of the viroses of plant
Microbial pesticide.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides one plant of nontoxic, no pathogenicities, to safety of human and livestock, to environment friend
It is good, the Bei Laisi bacillus of resistance is strong, bacteriostatic activity is stable inhibition plant virus, promotion plant growth, and answered
For prevent and treat the viroses of plant and fungal diseases of plants, promote plant growth etc., realizes and efficiently inhibit virus, promote to plant
Object growth, does not destroy the purpose of natural environment, and then realize sustainable, harmonious development.
One plant of inhibition is viral, promotes the entitled Bei Laisi bacillus of the Bei Laisi bacillus of plant growth
(Bacillus velezensis) SNB55, depositary institution are in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart, deposit number are CGMCC NO.17315, and the deposit date is on March 8th, 2019.Preservation address: the Chaoyang District, Beijing City North Star
No. 3 Institute of Microorganism, Academia Sinica, institute of West Road 1.
As a preferred solution, the bacterium of the described Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth
Strain cultural characteristic is that bacterium colony is white to milky on beef extract-peptone agar plate, and subcircular, coarse and edge is not advised
Then, opaque, there is a fold on surface, and quality is viscous, matter shape；After Liquid Culture, standing can form white film in the liquid surface,
Muddiness is uniform in the liquid, does not agglomerate.
It is further preferred that the strain morphology of the Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth
Feature is that thallus is in the shape of a rod, and size is (0.5~0.6) μ m (1.5~3.2) μm, is condensed together in short chain or beads shape row
Column, amphitrichous can move；Gram-positive；Gemma ellipse, middle life, sporangiocyst expand, no parasporal crystal.
It is further preferred that the optimum of the Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth is raw
Long temperature is 26~28 DEG C.
It is further preferred that the tunning of the Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth
It is able to suppress plant virus and phytopathogen；The plant virus and phytopathogen include tobacco mosaic virus (TMV), tomato yellow
Change curve leaf disease virus, Bacillus cercus, cucumber fusarium axysporum, Tobacco target spot bacterium, phytophthora blight of pepper, Rhizoctonia solani Kuhn and kind
Solanum cinerea bacterium.
It is further preferred that described inhibit virus, promote the Bei Laisi bacillus SNB55 of plant growth that can secrete Yin
Indolylbutyric acid promotes seed sprouting and plant growth.
It is further preferred that the Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth and its fermentation produce
Object is nontoxic, no pathogenicity, and to safety of human and livestock, environmentally friendly, resistance is strong, bacteriostatic activity is stable.
One plant inhibits virus, promotes the Bei Laisi bacillus SNB55 of plant growth in the prevention and treatment viroses of plant and plant
Application in fungal disease.
As a preferred solution, described to inhibit virus, the Bei Laisi bacillus SNB55 of plant growth is promoted to promote
Into the application in plant growth.
It is further preferred that the Bei Laisi bacillus SNB55 for inhibiting virus, promoting plant growth is applied to prevention and treatment
Tobacco mosaic virus disease, tomato yellow leaf curl virus disease, cucumber fusarium axysporum, Tobacco target spot, capsicum epidemic disease, rice sheath blight disease and
Graw mold of tomato.
The beneficial effects of the present invention are: the one plant provided by the invention Bei Laisi bud for inhibiting virus, promoting plant growth
Spore bacillus and its application have the advantage that
1. provide one plant inhibit virus, the Bei Laisi bacillus SNB55 for promoting plant growth and its tunning without
Poison, no pathogenicity, to safety of human and livestock, environmentally friendly, it is one plant of multi-functional biocontrol microorganisms that resistance is strong, bacteriostatic activity is stables, is had
Have broad application prospects.
2. one plant the provided Bei Laisi bacillus SNB55 and its tunning energy for inhibiting virus, promoting plant growth
The inhibition plant virus of enough wide spectrums and phytopathogen, including tobacco mosaic virus (TMV), tomato yellow leaf curl virus, wax-like bud
Spore bacillus, cucumber fusarium axysporum, Tobacco target spot bacterium, phytophthora blight of pepper, Rhizoctonia solani Kuhn and botrytis cinerea.Especially
Stronger inhibiting effect is all had to tobacco mosaic virus (TMV) and tomato yellow leaf curl virus, its fermentation liquid is prepared into biological and ecological methods to prevent plant disease, pests, and erosion water
Agent, Seedling Stage sprays, pouring root, capsuled seed have tomato virus disease caused by tobacco mosaic virus (TMV) and tomato yellow leaf curl virus
There is good control efficiency.
3. one plant provided inhibits virus, promotes the Bei Laisi bacillus SNB55 of plant growth that can secrete heteroauxin
(IAA), tomato seeds are promoted to germinate, take root and growth of seedling.
4. one plant provided inhibits virus, the Bei Laisi bacillus SNB55 of plant growth is promoted to increase prevention and treatment plant
The type of the microorganism antivirotic of virosis.
Detailed description of the invention
Fig. 1 is that one plant of inhibition of the invention is viral, promotes the colonial morphology of the Bei Laisi bacillus SNB55 of plant growth
Figure；
Fig. 2 is one plant of Bei Laisi bacillus SNB55 fermentation liquid and hair for inhibiting virus, promoting plant growth of the invention
Antibacterial loop graph of the ferment filtrate to Bacillus cercus；
Fig. 3 is that pot experiment measures ferment filtrate on Nicotiana glutinosa to the inhibiting effect test of TMV in the embodiment of the present invention
The control group Nicotiana glutinosa of middle inoculation TMV buffer；
Fig. 4 is that pot experiment measures ferment filtrate on Nicotiana glutinosa to the inhibiting effect test of TMV in the embodiment of the present invention
Middle SNB55 ferment filtrate is inoculated with Nicotiana glutinosa after mixing with 2 times of TMV buffers；
Fig. 5 is control group 2 in ferment filtrate influences tomato seeds and growth of seedling in the embodiment of the present invention detection
Seed is taken root situation；
Fig. 6 is experimental group 2 in ferment filtrate influences tomato seeds and growth of seedling in the embodiment of the present invention detection
Seed is taken root situation；
Fig. 7 is control group 3 in ferment filtrate influences tomato seeds and growth of seedling in the embodiment of the present invention detection
Growth of seedling situation；
Fig. 8 is experimental group 3 in ferment filtrate influences tomato seeds and growth of seedling in the embodiment of the present invention detection
Growth of seedling situation.
Appended drawing reference: 1- fermentation liquid inhibition zone, 2- ferment filtrate inhibition zone.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
Every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
One plant of Bei Laisi bacillus for inhibiting virus, promoting plant growth, the strain name are Bei Laisi bacillus
(Bacillus velezensis) SNB55, depositary institution are in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart, deposit number are CGMCC NO.17315, and the deposit date is on March 8th, 2019.Preservation address: the Chaoyang District, Beijing City North Star
No. 3 Institute of Microorganism, Academia Sinica, institute of West Road 1.
1. strain culturing
(1) culture medium forms
Plate, slant medium be beef extract-peptone agar medium: beef extract 3g, peptone 5g, sodium chloride 5g,
Agar 20g adds distilled water to 1000mL, pH 7.0~7.2.
Fermentation medium: beef extract 3g, peptone 5g, sodium chloride 5g add distilled water to 1000mL, pH 7.0~7.2.
Bacillus cercus culture medium: beef extract 3g, peptone 3g, Na2HPO40.1g, 18~20g of agar add distillation
Water is to 1000mL, pH 7.8~8.0.
(2) Bei Laisi bacillus SNB55 culture and ferment filtrate preparation
Inclined-plane culture: bacterial strain SNB55 being forwarded on fresh slant medium, is cultivated 1d under the conditions of 28 DEG C, is obtained into
Ripe slant pore；
Fermented and cultured: aseptically, aforementioned mature slant pore is seeded to 5% inoculum concentration and is fermented equipped with 50mL
In the 250mL triangular flask of culture medium, the shaken cultivation 46h on 28 DEG C, the rotary shaker of revolving speed 150r/min obtains fermentation liquid；
Sterile ferment filtrate preparation: being centrifuged 20min for aforesaid fermentation liquid under conditions of 10000r/min, collect supernatant,
WithBiofilter filtering, obtain sterile ferment filtrate.
2. bacterial strain detects
(1) detection project and method
1) colonial morphology detects: by bacterial strain SNB55, sectional streak, 28 DEG C of culture 5d observe single bacterium on plating medium
Fall form.
2) bacterial strain SNB55 inhibits Bacillus cercus (Bacillus cereus) Activity determination: will contain 109cfu/mL
Bacillus cercus 700 μ L of bacteria suspension mixed with the Bacillus cercus culture medium that 70mL melts after pour into square plate and be made
200 μ L aforesaid fermentation liquid are added in plate in an Oxford cup, and the aforementioned sterile fermentation filter of 200 μ L is added in another Oxford cup
Liquid measures antibacterial circle diameter after 28 DEG C of culture 16h.
3) pot experiment measures ferment filtrate on Nicotiana glutinosa to the inhibiting effect of TMV: using juice frictional inoculation method, people
Work is inoculated with TMV in measuring ferment filtrate on the healthy Nicotiana glutinosa of potting to the inhibiting effect of TMV, and experiment is divided into 4 experimental groups and 1
A control group, is repeated 3 times by every group 30 plants, and specific processing method see the table below 1.Withered spot number is investigated after inoculation 5d, scab is calculated and inhibits
Rate.
1 pot experiment of table measures ferment filtrate on Nicotiana glutinosa to the inhibiting effect experimental group processing method of TMV
4) pot experiment measures ferment filtrate on tomato to the inhibiting effect of TMV: tomato variety L402, using juice
Frictional inoculation method, artificial infection TMV measure ferment filtrate to the inhibitory activity of TMV, experiment point in the healthy tomato leaf of potting
For 3 experimental groups and 1 control group, every group 30 plants, it is repeated 3 times, specific processing method see the table below 2.Distinguished according to grade scale
Tomato incidence is recorded, calculates control efficiency, and be administered laggard row crop security survey.
2 pot experiment of table measures ferment filtrate on tomato to the inhibiting effect experimental group processing method of TMV
Grade scale: 0 grade: without any symptom；1 grade: the vein of lobus cardiacus is bright arteries and veins, and floral leaf is presented in 1-2 piece true leaf；3 grades: in
Upper blade floral leaf；5 grades: most blade floral leaves, a small number of leaf malformations；7 grades: most blades are attached most importance to floral leaf, deformity, shrinkage；9
Grade: nearly all blade is attached most importance to floral leaf, deformity, shrinkage, and plant is downgraded more apparent.
5) preventive effect of the pot experiment measurement ferment filtrate to Tomato yellow mosaic virus: tomato variety L402 sets 5 altogether
Group, including 3 experimental groups and 2 control groups.Experimental group 1: tomato temporary planting 3d starts to spray the ferment filtrate of 20 times of dilution, interval
5d is sprayed 1 time again, is sprayed 3 times altogether；Experimental group 2: impregnating seed 5h with the ferment filtrate for diluting 20 times, normal to plant；Experimental group 3:
With the ferment filtrate pouring root for diluting 20 times after tomato temporary planting 3d, every plant of 10mL, interval 5d are filled 1 time again, are filled 3 times altogether；Control group 1:
Spray clear water；Control group 2: spray amino-oligosaccharide (island element).Using juice frictional inoculation method, connect in 5 leaf period frictions of tomato
Kind TMV, 20d investigates diseased plant incidence according to tomato TMV disease symptoms grade scale after inoculation, calculates each processing disease index
And control efficiency.
Tomato TMV disease symptoms grade scale: 0 grade: without any symptom；1 grade: the vein of lobus cardiacus is bright arteries and veins, and 1-2 piece true leaf is in
Existing floral leaf；3 grades: middle and upper part blade floral leaf；5 grades: most blade floral leaves, a small number of leaf malformations；7 grades: most blades attach most importance to floral leaf,
Deformity, shrinkage；9 grades: nearly all blade is attached most importance to floral leaf, deformity, shrinkage, and plant is downgraded more apparent.
6) preventive effect of the pot experiment measurement ferment filtrate to tomato yellow leaf curl virus disease: tomato variety L402 sets 3 altogether
A group, including 1 experimental group and 2 control groups.Experimental group 1: starting to spray the ferment filtrate of 20 times of dilution in tomato temporary planting 3d,
Interval 5d sprays 1 time again, sprays 3 times altogether；Control group 1: spray clear water；Control group 2: spray amino-oligosaccharide (island element).It is administered at the 3rd time
The aleyrodid with tomato yellow leaf curl virus that malicious source chamber is raised uniformly is discharged on test tomato plant afterwards, carries out passing poison,
Guarantee on every plant of tomato 10 or more, diseased plant is investigated according to tomato yellow leaf curl virus disease symptoms grade scale in 15d after the onset
Incidence calculates diseased plant rate, disease index and control efficiency.
3 tomato yellow leaf curl virus disease symptoms grade scale of table
7) preventive effect of the field plot trial measurement ferment filtrate to tomato yellow leaf curl virus disease: tomato variety L402,
Fly net is arranged to obstruct, sets 5 groups, including 3 experimental groups and 2 control groups, every group of 3 repetitions altogether.Experimental group 1: tomato field planting
The ferment filtrate of 20 times of dilution is sprayed after 3d, interval 5d is sprayed 1 time again, sprayed 3 times altogether；Experimental group 2: with the ferment filtrate for diluting 20 times
Seed 5h is impregnated, it is normal to cultivate；Experimental group 3: tomato is colonized after 3d with 20 times of ferment filtrate pouring root of dilution, every plant of 10mL,
It fills 1 time every 5d, fills 3 times altogether；Control group 1: spray clear water；Control group 2: spray amino-oligosaccharide (island element).After processing 10d into canopy
The aleyrodid with TYLCV is discharged, whole process does not spray any insecticide, and 15d is by tomato yellow leaf curl virus disease symptoms classification mark after the onset
Quasi- investigation, calculates disease index and control efficiency.
8) detection that ferment filtrate influences tomato seeds and growth of seedling: experiment sets 6 groups, including 3 experimental groups and 3 altogether
A control group, every group of 3 repetitions；Wherein experimental group 1, experimental group 2 and control group 1, control group 2 are used to influence seed growth
Detection；The detection of experimental group 3 and control group 3 for the influence to growth of seedling.
1. the detection influenced on seed growth
Experimental group 1: ferment filtrate impregnates tomato seeds 5h, and seed is put into the sterile petri dish for being covered with wet filter paper later
In, 28 DEG C of heat insulating cultures observe seed sprouting situation；Experimental group 2: the ferment filtrate of 20 times of dilution impregnates tomato seeds 5h, it
Seed is put into afterwards in the sterile petri dish for being covered with wet filter paper, 28 DEG C of heat insulating cultures, observes seed sprouting situation；Control group 1:
Fermentation medium impregnates tomato seeds 5h, is put into seed in the sterile petri dish for being covered with wet filter paper later, 28 DEG C of heat preservation trainings
It supports, observes seed sprouting situation；Control group 2: the fermentation medium of 20 times of dilution impregnates tomato seeds 5h, is later put into seed
It is covered in the sterile petri dish of wet filter paper, 28 DEG C of heat insulating cultures, observes seed sprouting situation.
2. the detection of the influence to growth of seedling
Experimental group 3: the ferment filtrate of 20 times of dilution impregnates tomato seeds 5h, and the vernalization in 28 DEG C of insulating boxs is sent out to seed
It after bud, is seeded in culturing pot, is placed in a greenhouse normal management；Control group 3: the fermentation medium of 20 times of dilution impregnates tomato species
Sub- 5h, vernalization is seeded in culturing pot after germination in 28 DEG C of insulating boxs, is placed in a greenhouse normal management.One month
After lift seedlings, clear water is cleaned blotted with blotting paper after measure plant height, root, overground part fresh weight and underground part fresh weight；It, will after weighing fresh weight
Tomato is placed in aeration cabinet 105 DEG C of water-removing 30min, and 75 DEG C weigh overground part and underground part dry weight after drying to constant weight.
9) the broad-spectrum antibacterial test of SNB55: taking diameter is Rhizoctonia solani Kuhn, cucumber fusarium axysporum, the tobacco target spot of 5mm
Germ, the back-off of phytophthora blight of pepper dish are in PDA plate center, at the bacterium dish up and down about 2cm, with speckling with Bei Laisi gemma bar
The oese of bacterium SNB55 draws two parallel lines, and 28 DEG C of culture 3d observe antibacterial situation.
10) SNB55 secretes the measurement of heteroauxin (IAA): the preparation of experimental group sample, by Bei Laisi bacillus
SNB55 is inoculated in containing L-Trp (200mgL-1) LB liquid medium in, 28 DEG C, 180rpm shaking table culture 4d.Take 50
μ L bacteria suspension drop adds 50 μ L SaLkowski color solution (50mL 35%HCLO on whiteware plate4+1mL 0.5M
FeCL3)(Libbert&Risch,1969).50 μ L (50mgL of IAA is added-1, Shanghai Sheng Gong bioengineering Co., Ltd,
Purity > 99%) titer be control group.The whiteware plate of experimental group and control group is seen after room temperature avoid light place 30min
It examines, the color person of reddening indicates being capable of producing IAA.
(2) testing result
1) colonial morphology testing result: the bacterial strain SNB55 bacterium colony on plating medium is white to milky, nearly circle
Shape, coarse and edge is irregular, opaque, and there is fold on surface, and quality cement shape is shown in Fig. 1；After Liquid Culture, standing can be in liquid
Body surface face forms white film, and muddiness uniformly, is not agglomerated in liquid, and strain morphology feature is that thallus is in the shape of a rod, and size is (0.5
~0.6) μ m (1.5~3.2) μm, condensing together can move in short chain or the arrangement of beads shape, amphitrichous；Gram's staining
It is positive；Gemma ellipse, middle life, sporangiocyst expand, no parasporal crystal.
2) bacterial strain SNB55 inhibits Bacillus cercus Activity determination result: the fermentation liquid of bacterial strain SNB55 and sterile fermentation filter
Liquid all has very strong inhibiting effect to Bacillus cercus, and concrete outcome is shown in Table 4, and inhibition zone situation is shown in Fig. 2.
The inhibitory effect of 4 bacterial strain SNB55 fermentation liquid of table and sterile ferment filtrate to Bacillus cercus
As can be seen from the table, the processing antibacterial circle diameter of fermentation liquid reaches 22mm, and inhibition zone is clear, transparent, and room temperature is put
It is observed after setting 15d, inhibition zone is still clear, transparent, and size is unchanged, illustrates the fermentation liquid of bacterial strain SNB55 to Bacillus cercus
Inhibiting effect is strong, and fungistatic effect is lasting.Fermentation liquid and sterile ferment filtrate to the antibacterial circle diameter size of Bacillus cercus without
Notable difference illustrates that Substance is mainly the tunning of bacterial strain SNB55.
3) pot experiment measures ferment filtrate on Nicotiana glutinosa to the inhibiting effect test result of TMV: bacterial strain SNB55 fermentation
The various processing of filtrate can inhibit infecting for TMV, reduce withered spot quantity on Nicotiana glutinosa, concrete outcome is shown in Table 5, and passivation is shown in figure
3 and Fig. 4.
5 bacterial strain SNB55 ferment filtrate of table is on Nicotiana glutinosa to the inhibiting effect of TMV
As can be seen from Table 5, the processing that ferment filtrate is mixed with TMV is most strong to the passivation of TMV, and withered spot inhibits
Rate is up to 94.75%；The inhibiting rate that 48h is sprayed before inoculation is 84.99%, and the inhibiting rate sprayed for 24 hours before inoculation is
80.70%；The inhibiting rate sprayed for 24 hours after inoculation is 70.18%.The withered spot inhibiting rate sprayed before inoculation sprays after being apparently higher than inoculation
The inhibiting rate to TMV is applied, illustrates that prevention effect is better than therapeutic effect, and 48h spraying effect is more preferable in advance.
4) pot experiment measures ferment filtrate on tomato to the inhibiting effect test result of TMV: bacterial strain SNB55 fermentation filter
The various processing of liquid have certain inhibiting effect to TMV, and concrete outcome is shown in Table 6.
6 bacterial strain SNB55 ferment filtrate of table is on tomato to the inhibiting effect of TMV
As can be seen from the table, inhibiting virus activity strongest is processing that ferment filtrate is directly mixed with TMV, fermentation filter
Liquid dilutes 20 times of liquid to the preventive effect of TMV up to 80%；The prevention effect sprayed before inoculation is 62.40%, hence it is evident that after being higher than inoculation
The therapeutic effect (50.71%) sprayed, and the growth rate of ferment filtrate processing disease incidence is also relatively slow, tomato growing way is good,
Color is green, vane extension, plumpness.
5) pot experiment measurement ferment filtrate is to the efficiency test result of Tomato yellow mosaic virus: experimental group 1, experimental group 2,
The disease index of tomato significantly reduces in experimental group 3, and preventive effect is respectively 63.25%, 61.80% and 66.51%, is above control
Group 2, root irrigation preventive effect > foliage-spray preventive effect > seed pelleting preventive effect, concrete outcome is shown in Table 7.And kind handled with ferment filtrate
Eggplant growing way is good, and color is green, and stalk is more sturdy, and the tomato plant of control group is short and small, yellow leaf.
Preventive effect of the 7 bacterial strain SNB55 ferment filtrate of table to Tomato yellow mosaic virus
3) efficiency test result of the pot experiment measurement ferment filtrate to tomato yellow leaf curl virus disease: SNB55 fermentation filter
Liquid can inhibit tomato yellow leaf curl virus to infect, to reduce tomato morbidity, concrete outcome is shown in Table 8.
Preventive effect of the 8 bacterial strain SNB55 ferment filtrate of table to tomato yellow leaf curl virus disease
As can be seen from the table, it is inoculated with before TYLCV with 20 times of dilution of ferment filtrate processing to tomato yellow leaf curl virus disease
Preventive effect be 56.30%, the preventive effect of control group 2 is 43.20%, and can significantly reduce disease incidence.
4) efficiency test result of the field plot trial measurement ferment filtrate to tomato yellow leaf curl virus disease: bacterial strain
SNB55 ferment filtrate has preferable control efficiency to tomato yellow leaf curl virus disease, and is better than amino-oligosaccharide, concrete outcome
It is shown in Table 9.
9 bacterial strain SNB55 ferment filtrate of table is to tomato yellow leaf curl virus disease field plot preventive effect
35d or so starts to fall ill after tomato field planting, and 15d is investigated after the onset, as can be seen from Table 9, experimental group 1, experiment
The disease index of the tomato of group 2, experimental group 3 significantly reduces, and preventive effect is respectively 52.86%, 31.91%, 56.31%, experimental group
Preventive effect be higher than medicament control group 2, and root irrigation preventive effect > foliage-spray preventive effect > seed pelleting preventive effect sprays bacterial strain SNB55
The experimental group of ferment filtrate is obviously fallen ill relatively gently, and plant growing way is high.
5) testing result that ferment filtrate influences tomato seeds and growth of seedling:
1. the influence to seed growth: SNB55 ferment filtrate stoste can inhibit seed to sprout, bacterial strain SNB55 ferment filtrate
Tomato Seeds Germination, hestening rooting can be promoted by diluting 20 times of liquid, and concrete outcome is shown in Table 10, and seed situation of taking root is shown in Fig. 5 and Fig. 6.
10 ferment filtrate of table is to tomato seeds growth effect testing result
As can be seen from the table, the seed of bacterial strain SNB55 ferment filtrate stoste processing is shorter by 42.86% than compareing, and ferments
The seed that filtrate dilutes 20 times of processing is sprouted in advance, is taken root in advance than control, root system increases, when the root long of fermentation dilution processing
When reaching 5cm, increase by 42.86% than control.
2. the influence to growth of seedling: 20 times of dilutions of bacterial strain SNB55 ferment filtrate have the function of promotion growth of seedling,
Concrete outcome is shown in Table 11, and growth of seedling situation is shown in Fig. 7 and Fig. 8.
Growth promoting function of the 11 bacterial strain SNB55 fermentation liquid of table to tomato seedling
The seedling grown up to as can be seen from the table with the tomato seeds of 20 times of dilution of SNB55 ferment filtrate processing, growing way
Good, color is dark green, and no matter the indexs and growing way etc. such as plant height, root long, overground part fresh weight/dry weight, underground part fresh weight/dry weight are obvious
Better than control, increase by 12.07%, 8.53%, 23.77%, 31.88%, 16.22%, 34.51% than control respectively.
6) the broad-spectrum antibacterial test result of SNB55: SNB55 is to cucumber fusarium axysporum, Tobacco target spot bacterium, Phytophthora capsici
Bacterium, botrytis cinerea and Rhizoctonia solani Kuhn all have certain inhibiting effect, wherein to Tobacco target spot bacterium inhibiting effect
It is most strong, followed by Rhizoctonia solani Kuhn.
7) SNB55 secretes the measurement result of heteroauxin (IAA): SaLkowski color solution is added in bacterial strain SNB55 bacteria suspension
Color reddens color afterwards, shows that bacterial strain SNB55 can producing IAA.
It should be appreciated that described above, the specific embodiments are only for explaining the present invention, is not intended to limit the present invention.By
The obvious changes or variations that spirit of the invention is extended out are still in the protection scope of this invention.
Claims (10)
1. one plant of Bei Laisi bacillus for inhibiting virus, promoting plant growth, which is characterized in that the strain name is Bei Laisi
Bacillus (Bacillus velezensis) SNB55, depositary institution are that China Committee for Culture Collection of Microorganisms is common
Microorganism center, deposit number are CGMCC NO.17315, and the deposit date is on March 8th, 2019.
2. the one plant according to claim 1 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature exist
In the strain culturing feature of the Bei Laisi bacillus SNB55 is that bacterium colony is white on beef extract-peptone agar plate
To milky, subcircular, coarse and edge is irregular, opaque, and there is a fold on surface, and quality is viscous, matter shape；It is quiet after Liquid Culture
White film can be formed in liquid surface by setting, in the liquid it is muddy uniformly, do not agglomerate.
3. the one plant according to claim 1 or 2 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature
It is, the strain morphology feature of the Bei Laisi bacillus SNB55 is that thallus is in the shape of a rod, and size is (0.5~0.6) μ m
(1.5~3.2) μm, condensing together can move in short chain or the arrangement of beads shape, amphitrichous；Gram-positive；Gemma is ellipse
Circle, middle life, sporangiocyst expand, no parasporal crystal.
4. the one plant according to claim 3 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature exist
In the optimum growth temperature of the Bei Laisi bacillus SNB55 is 26~28 DEG C.
5. the one plant according to claim 4 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature exist
In the tunning of the Bei Laisi bacillus SNB55 is able to suppress plant virus and phytopathogen；The plant virus
It include tobacco mosaic virus (TMV), tomato yellow leaf curl virus, Bacillus cercus, cucumber fusarium axysporum, tobacco with phytopathogen
Target spot germ, phytophthora blight of pepper, Rhizoctonia solani Kuhn and botrytis cinerea.
6. the one plant according to claim 5 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature exist
In the Bei Laisi bacillus SNB55 can secrete heteroauxin, promote seed sprouting and plant growth.
7. the one plant according to claim 5 Bei Laisi bacillus for inhibiting virus, promoting plant growth, feature exist
The nontoxic, no pathogenicity in, the Bei Laisi bacillus SNB55 and its tunning resists safety of human and livestock, environmentally friendly
Inverse property is strong, bacteriostatic activity is stablized.
8. the application of the Bei Laisi bacillus of one plant of inhibition virus, promotion plant growth, which is characterized in that the Bei Laisi bud
Application of the spore bacillus SNB55 in the prevention and treatment viroses of plant and fungal diseases of plants.
9. one plant of inhibition is viral according to claim 8, promotes the application of the Bei Laisi bacillus of plant growth, feature
It is, the Bei Laisi bacillus SNB55 is promoting the application in plant growth.
10. one plant of inhibition is viral according to claim 9, promotes the application of the Bei Laisi bacillus of plant growth, special
Sign is that the Bei Laisi bacillus SNB55 is applied to prevention and treatment tobacco mosaic virus disease, tomato yellow leaf curl virus disease, Huang
Cucurbit wilt, Tobacco target spot, capsicum epidemic disease, rice sheath blight disease and graw mold of tomato.
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