CN106857040A - A kind of prevention and controls and composition for cape jasmine leaf blight disease - Google Patents
A kind of prevention and controls and composition for cape jasmine leaf blight disease Download PDFInfo
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- CN106857040A CN106857040A CN201611231643.2A CN201611231643A CN106857040A CN 106857040 A CN106857040 A CN 106857040A CN 201611231643 A CN201611231643 A CN 201611231643A CN 106857040 A CN106857040 A CN 106857040A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/64—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with three nitrogen atoms as the only ring hetero atoms
- A01N43/647—Triazoles; Hydrogenated triazoles
- A01N43/653—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/08—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having one or more single bonds to nitrogen atoms
- A01N47/10—Carbamic acid derivatives, i.e. containing the group —O—CO—N<; Thio analogues thereof
- A01N47/18—Carbamic acid derivatives, i.e. containing the group —O—CO—N<; Thio analogues thereof containing a —O—CO—N< group, or a thio analogue thereof, directly attached to a heterocyclic or cycloaliphatic ring
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Abstract
The invention provides a kind of prevention and controls and composition for cape jasmine leaf blight disease, the technical scheme is primarily based on Koch's Postulates, combining form observation, microscopical characters and the analysis of fungi rDNA ITS sequences carry out the symptom and pathogen identification of system to cape jasmine leaf blight pathogen, give clear and definite analytical conclusions to the pathogenic microorganism for causing cape jasmine leaf blight first.Experiment finds that leaf blight result of study shows：Causing the pathogen of cape jasmine leaf blight disease has three kinds, respectively Phomopsis (Phomopsis sp.), Alternaria tenuissima (Alternaria tenuissima (Fr.) Wiltshire) and alternaric bacteria (Alternaria alternata (Fr.) Keissler).Based on more than beneficial discovery, present invention determine that a kind of to kill above-mentioned three kinds of microorganisms as the cape jasmine leaf blight pest control method of core.On this basis, the present invention by experiment in vitro has investigated fungistatic effect of the different bacteriostatic agents to above-mentioned three kinds of microorganisms, so that it is determined that for the composition of above-mentioned cape jasmine leaf blight pest control method.
Description
Technical field
The present invention relates to plant disease technical field, further to probing into and drug sieve for plant disease mechanism
A kind of choosing, and in particular to prevention and controls and composition for cape jasmine leaf blight disease.
Background technology
Cape jasmine (Gardenia jasminoides Eills) is Rubiaceae (Rubiaceae) plant, and medicinal part is it
Dry mature fruit, containing Geniposide glycoside, crocin class isoreactivity composition, with liver protection, cholagogic, calmness, is depressured, disappears
The effects such as scorching, hemostasis.The total glycosides of west safflower in cape jasmine fruit can be used to produce gardenia red, Gardenia Yellow, three kinds of pigments of gardenia blue,
Widely used in food service industry as natural colorant.Because cape jasmine fruit is widely used at the aspect such as medicine and food industry,
It is very big to the demand of cape jasmine resource both at home and abroad.
In the cultivation of cape jasmine, defect phenomenon is one of accident the most serious to yield effect, existing skill
In art the multiple disease of cape jasmine mainly including leaf blight, samping off, root rot, root disease etc. is macerated, wherein again with leaf blight influence most
For serious.Symptoms aspect, leaf blight is more to infect generation from leaf margin, blade tip, and scab is ascending irregular, and bronzing is extremely
Taupe, in flakes into big withered spot, dry up scab area up to the 1/3-1/2 of blade, and there is a band deep compared with scab at scab edge；Disease is strong
Boundary is obvious.Later stage produces some black small grain points on scab.The Activities of Some Plants disease leaf initial stage first turns yellow, and yl moiety gradually becomes
Brown necrosis.By extending partially into whole vein, be presented brown to leaf margin scab russet, scab marginal wavy, color compared with
It is deep.Sick key has a common boundary substantially, and its outer rim also has the shallow band of yellow that width is not waited sometimes, and then, scab gradually extends to leaf base, directly
It is changed into brown to taupe to whole blade.Then there is black villiform thing or black dot in sick leaf back or front.
Although leaf blight is widely present in various plants, the intercommunity of different plant leaf blights is only limitted to Symptoms
And disease name, and often there is basic sex differernce in pathogenesis aspect.In this case, cape jasmine leaf blight should be directed to
Actual characteristic study its pathogenesis, pesticide control screening is targetedly carried out in the case of clear and definite pathogenic microorganisms,
The prevention effect that may have been obtained.The report to cape jasmine leaf blight pest control method is had no in the prior art.
The content of the invention
It is contemplated that for the technological deficiency of prior art, there is provided a kind of prevention and controls for cape jasmine leaf blight disease
And composition, to solve the indefinite technical problem of cape jasmine leaf blight cause of disease in the prior art.
Another technical problem to be solved by the present invention is that the prevention effect of cape jasmine leaf blight is not good in the prior art.
To realize above technical purpose, the present invention uses following technical scheme：
A kind of prevention and controls for cape jasmine leaf blight disease, the method is antibacterial to cape jasmine plant applying Phomopsis
Agent, Alternaria tenuissima bacteriostatic agent, alternaric bacteria bacteriostatic agent.
Preferably, the Phomopsis bacteriostatic agent is N- (2- benzimidazoles base)-methyl carbamate；Further preferably
, the concentration of N- (2- benzimidazoles base)-methyl carbamate is 0.19 μ g/mL.
Preferably, the Alternaria tenuissima bacteriostatic agent is hexaconazole；It is further preferred that the concentration of hexaconazole is 9.33
μg/mL。
Preferably, the alternaric bacteria bacteriostatic agent is hexaconazole；It is further preferred that the concentration of hexaconazole is 14.45 μ
g/mL。
Meanwhile, the invention provides a kind of composition for the above method, said composition includes N- (2- benzimidazoles
Base)-methyl carbamate and hexaconazole.
Preferably, the concentration of N- (2- benzimidazoles base)-methyl carbamate is 15 μ gmL in said composition-1。
Preferably, the concentration of hexaconazole is 500 μ gmL in said composition-1。
In above technical scheme, the bacteriostatic agent is not limited to play growth of microorganism situation the thing of inhibitory action
Matter, all there is the material of growth inhibition effect or direct killing action all should work as to prescribed microorganism belong to the bacteriostatic agent institute
The technical scope of restriction.The bacteriostatic agent should understand to define its implication with the generality of those skilled in the art.
The invention provides a kind of prevention and controls and composition for cape jasmine leaf blight disease, technical scheme base first
In Koch's Postulates, combining form observation, microscopical characters and fungi rDNA-ITS sequence analyses are to cape jasmine leaf blight pathogen
The symptom and pathogen identification of the system of carrying out, give clearly analysis knot to the pathogenic microorganism for causing cape jasmine leaf blight first
By.Experiment finds that leaf blight result of study shows：Causing the pathogen of cape jasmine leaf blight disease has three kinds, respectively intends stem point
Mould (Phomopsis sp.), Alternaria tenuissima (Alternaria tenuissima (Fr.) Wiltshire) and alternaric bacteria
(Alternaria alternata(Fr.)Keissler).The beneficial discovery based on more than, present invention determine that a kind of killing
Above-mentioned three kinds of microorganisms are the cape jasmine leaf blight pest control method of core.
On this basis, the present invention has investigated antibacterial effect of the different bacteriostatic agents to above-mentioned three kinds of microorganisms by experiment in vitro
Really.PDA plate bacteriostatic experiment result shows：N- (2- benzimidazoles base)-methyl carbamate (carbendazim) is to Phomopsis
Bactericidal effect is best, EC50It is 0.19 μ gmL-1, secondly it is five nitre carbendazim, thiophanate-methyl, thiophanate methyl, Bravo；
And the optimal Fungicidal substance for being directed to Alternaria tenuissima and alternaric bacteria is hexaconazole, the medium effective concentration difference of the two is as little as
9.33 μ g/mL and 14.45 μ g/mL.Under the guide of above Experiment Result, present invention determine that for above-mentioned cape jasmine leaf blight disease
The composition of evil prevention and controls, so that for the exploitation of cape jasmine leaf blight disease control medicine is laid a good foundation.The inventive method with
Based on the discovery of cape jasmine leaf blight pathogenic microorganisms, the specific aim of means of prevention is improved, cape jasmine leaf blight has been effectively ensured
Disease-controlling effect.
Brief description of the drawings
Fig. 1 is isolated GD1 bacterial strain colonial morphology figures in the embodiment of the present invention 1；
Fig. 2 is isolated GD2 bacterial strain colonial morphology figures in the embodiment of the present invention 1；
Fig. 3 is isolated GD3 bacterial strain colonial morphology figures in the embodiment of the present invention 1；
Fig. 4 is isolated GD1 bacterial strain spore microscopy figures in the embodiment of the present invention 1；
Fig. 5 is isolated GD1 bacterial strain mycelium microscopy figures in the embodiment of the present invention 1；
Fig. 6 is isolated GD2 bacterial strain mycelium microscopy figures in the embodiment of the present invention 1；
Fig. 7 is isolated GD3 bacterial strain mycelium microscopy figures in the embodiment of the present invention 1；
Fig. 8 is by pathogen bacterium isolated again from blade after GD1 inoculations to blade in the embodiment of the present invention 1
Fall aspect graph；
Fig. 9 is by pathogen bacterium isolated again from blade after GD2 inoculations to blade in the embodiment of the present invention 1
Fall aspect graph；
Figure 10 is by pathogen isolated again from blade after GD3 inoculations to blade in the embodiment of the present invention 1
Colonial morphology figure；
Figure 11 is the disease incidence figure that cape jasmine blade after GD1 bacterial strains is inoculated with the embodiment of the present invention 1；
Figure 12 is the disease incidence figure that cape jasmine blade after GD2 bacterial strains is inoculated with the embodiment of the present invention 1；
Figure 13 is the disease incidence figure that cape jasmine blade after GD3 bacterial strains is inoculated with the embodiment of the present invention 1；
Figure 14 is the rDNA-ITS areas PCR amplifications of GD1 bacterial strains in the embodiment of the present invention 1；
Figure 15 is the rDNA-ITS areas PCR amplifications of GD2 bacterial strains in the embodiment of the present invention 1；
Figure 16 is the rDNA-ITS areas PCR amplifications of GD3 bacterial strains in the embodiment of the present invention 1.
Specific embodiment
Specific embodiment of the invention will be below described in detail.In order to avoid excessive unnecessary details,
Be will not be described in detail to belonging to known structure or function in following examples.
Approximating language used in following examples can be used for quantitative expression, show do not changing the feelings of basic function
Quantity can be allowed under condition certain variation.Therefore, it is accurate that the numerical value corrected with the language such as " about ", " left and right " is not limited to this
Numerical value is in itself.In certain embodiments, scope of the numerical value for allowing it to correct positive and negative 10 (10%) " about " is represented
Interior change, such as, what " about 100 " represented can be any numerical value between 90 to 110.Additionally, " the about first numerical value is arrived
In the statement of second value ", the first and second numerical value two values are at about corrected.In some cases, approximating language
May be relevant with the precision of measuring instrument.
In addition to being defined, technology used and scientific terminology have and art technology people of the present invention in following examples
The identical meanings that member is commonly understood by.Term used " first ", " second " etc. are not offered as any order, number in following examples
Amount or importance, and be only used for distinguishing a kind of element and another element.
Embodiment 1 (cape jasmine leaf blight pathogen identification)
1 test method
1.1 pathogenicbacteria separations
Cape jasmine leaf blight typical blade is selected, preliminary isolating and purifying is carried out to pathogen using conventional organization partition method.
Blade surface is rinsed well with distilled water.The tissue block of intersection clip 5mm5mm sizes is good in the disease of leaf spot lesion, in
30s is soaked in 70% ethanol, placing into solution immersion 5min in 5% sodium hypochlorite carries out surface sterilization, afterwards with aseptic
Washing goes residual solution, excess surface moisture to be blotted with aseptic blotting paper, be inoculated in PDA culture medium, and aforesaid operations are super
Carried out under aseptic condition on net workbench.Postvaccinal culture medium is placed in the dark training of 30 DEG C of LRH-250-5 constant temperature and humidities incubator
Support.Bacterium colony is initially formed after 2d is cultivated, and is beaten in edge diverse location and is taken mycelia, is transferred to culture in new PDA culture medium and is obtained
The bacterial strain that must be purified, purifying bacterial strain is transferred to slant medium, is saved backup in 4 DEG C.
1.2 pathogenic bacteria determine
Using Isolated leaf inoculation method.20, the healthy cape jasmine blade of maturation is adopted, with 75% ethanol by excised leaf surface
Carry out disinfection, sterilized water washes away residual ethanol, moisture volatilizes to be beaten after ripe bacterium colony surface and takes the bacteria cake of diameter 6mm and transfer in each
Blade surface, is put into porcelain dish, plus distilled water covering blade.30 DEG C of dark culturings in constant incubator.To be inoculated with blank training
The blade for supporting base is compared, and incidence is observed after 48h.Record the symptom and feature of disease, and Taking Pictures recording.
1.3 pathogens separate again
By the blade of the sequela of inoculation, same method during using pathogenicbacteria separation carries out separating again for pathogen.See
The morphological feature of the microorganism for recording isolated is examined, is verified with Koch's Postulates.
1.4 pathogen identifications
1.4.1 the morphologic observation of colonial morphology, mycelia and spore
Bacterium colony to different growing stage carries out morphological observation, and in different growth phases in light microscope (40
10 times) under observe mycelia and spore microscopic features, according to colonial morphology, Spore Types, asexual and generative propagation organ etc., reference
《Fungal identification handbook》、《Chinese fungi will》Etc. being identified.
1.4.2 Molecular Identification
Pathogen DNA is extracted using conventional CTAB methods：
The pathogenic strong and weak comparing of 1.5 pathogens
The pathogen of species will be had determined that carries out pharmacosensitive test, every kind of pathogen according to the method in 1.2 pathogenic bacteria measure
10 healthy leaveses are set, during incidence per 2h each blades of observed and recorded, it is determined that starting the time of morbidity, surveyed after 2d
Measure the lesion diameter of each pathogen infection blade, and Taking Pictures recording.
2 results and analysis
2.1 Disease symptoms investigation results
Leaf blight is Common mycotic disease, and whole year can occur.Early stage, blade tip or leaf margin occur oval or not
The patch of regular shape, in bronzing, as the state of an illness spreads development, scab is gradually extended from edge to leaf base, scab in flakes into
Big withered spot, dries up up to integrated plate blade, and there is a band deep compared with scab at scab edge, and the strong boundary of disease is obvious.In Ji'an Xiajiang cape jasmine
In planting base, cape jasmine leaf blight large area occurs, and lesion diameter causes plant fallen leaves, shedding or withered up to 2~5 centimetres,
Make whole strain withered when serious.
2.2 Morphological Characteristics of Their Pathogenic and pathogenic detection
2.2.1 Morphological Characteristics of Their Pathogenic
Isolated and purified by the cape jasmine leaf blight blade gathered to field, multiple fungal bacterial strains are obtained, according to Ke He
Family name's card disease rule carries out Pathogenicity, judges that wherein three bacterial strains are the sick pathogen, and GDl, GD2 are temporarily respectively designated as here
With GD3 (notes：GD--gardenia disease pathogen).
In PDA culture medium, bacterium colony is presented irregular petal-shaped radiation, white when mycelia children is tender, during culture 2d to bacterial strain GDl
Color is started from the center of bacterium colony and is progressively changed into light brown, rear colony colour is gradually deepened, 5-6d can cover with the flat of diameter 9cm
Plate, colonial morphology is shown in Fig. 1.Basis of microscopic observation hypha form, mycelia wall is thin, transparent, is grown in distorted shape, and width is irregular,
A diameter of 3-6 μm, there is obvious barrier film, microscopy figure is shown in Fig. 4, Fig. 5.Pycnidia is spherical to oblate spheroid, buries and is born in blade face, greatly
Small is 255~423 μm (n=30), wall dark brown, and substantially, conidiophore is colourless, separate, tufted or sympodial branching in aperture,
11~25 × 1.7~2.6 μm.There is α- conidia in pycnidia, colourless, spindle, unit cell includes l~3 oil
Ball, size is 5.14~10.8 × 1.9~4.4 μm (n=30).Features described above is consistent with the morphological feature of Phomopsis sp..
Bacterial strain GD2 grows vigorous in PDA culture medium, and bacterium colony is open and flat.Be at the beginning of bacterium colony white, after be changed into blackish green to black
Color.Bacterium colony is that suede is cotton-shaped, and aerial hyphae is flourishing, and mycelia has separation, multi-branched.The many fasciations of conidiophore, differentiation is obvious, Dan Sheng
Or fasciation, it is straight or slightly curved, it is filbert；There are separation, less branch.Conidium is concatenated, in nearly oval, the shape of falling club, surface
Smooth, a few surface spinosity wart, conidium has 3~8 tabulas, and 1~4 rake barrier film, separated place is not hung contracting, and size is
(23.7~41.5) μ m (9.2~15.8) μm, germ tube 1~3, multichain life, a small number of Dan Sheng.Short beak is in the form of a column or taper, portion
Divide and be changed into conidiogenous cell.Features described above is consistent with the morphological feature of Alternaria tenuissima (Fr.) Wiltshire.Bacterium
The form that falls is as shown in Fig. 2 microscope inspection figure is as shown in Figure 6.
Bacterial strain GD3 well-growns in PDA culture medium.Bacterium colony subcircular, cultivates in the culture dish of diameter 9cm at 30 DEG C
7d is to cover with full ware, and mycelium is intensive, villiform.It is white, later stage color burn, gradually in black at the beginning of mycelium.Microscope
The smooth tool of lower mycelia every, conidium is chain life, and base portion blunt circle, the other end is sharper, has and separates in length and breadth, general tabula 1~3,
Mediastinum 1~6, spore size is 5.9~12.5 μm of 9.9~35.2 μ m, and mean size is 7.9 μm of 18.8 μ m.Above-mentioned spy
Levy and be consistent with the morphological feature of Alternaria alternata (Fr.) Keissler.Colonial morphology is as shown in figure 3, microscope
Inspection figure is as shown in Figure 7.
2.2.2 pathogenic detection
Three kinds of tender leafs of pathogen mycelia moisturizing culture 2d at room temperature is inoculated with, is shown and abiogenous leaf blight phase
As symptom, and PDA culture medium inoculation blade do not show symptom.Taking the blade of inoculation morbidity, carry out can after pathogen separates again
Identical pathogen is obtained, GD1, GD2, GD3 is demonstrated and is pathogenic bacteria, and infected by blade need not be stabbed.Take and infect disease
The cape jasmine blade of opportunistic pathogen can obtain identical pathogen after carrying out pathogenicbacteria separation, as shown in figs. 8-10.Scab shows bronzing,
Slide measure measures 4.2~15.7mm of diameter.Subsequent scab expands, and color is progressively changed into dark brown or black, such as Figure 11~13
It is shown.
2.3 rDNA-ITS sections are expanded and are sequenced
RDNA-ITS sequence amplifications are carried out to three kinds of pathogens using pathogen template DNA and universal primer, is as a result obtained
The single band between 500-750bp is obtained, Figure 14~16 are seen.
2.4 homologys
Bacterial strain GD1 amplifications obtain the specific segment of 583bp, and sequencing result shows that homology highest bacterial strain is
Phomopsis sp. (Phomopsis bacterium), its similitude is 100%.Colonial morphology is consistent with microscopic features with it, therefore determines
The pathogen is Phomopsis (Phomopsis sp.).
Bacterial strain GD2 amplifications are the specific segment of 542bp, and sequencing result shows that homology bacterial strain higher is
Alternaria sp. (rod method), Alternaria alternata (Cross spectrum method) and Alternaria
Tenuissima (Alternaria tenuissima).Its similitude is up to 99%~100%.Form and micro- spy with reference to 2.2.1 bacterial strains GD2
Levy, determine that the pathogen is Alternaria tenuissima (Alternaria tenuissima (Fr.) Wiltshire).
The specific segment of 542bp is arrived in bacterial strain GD3 amplifications, and sequencing result shows that homology bacterial strain higher is
Alternaria sp. (rod method).Its similitude is 99%.Combining form feature determines that the bacterium is alternaric bacteria
(Alternaria alternata(Fr.)Keissler)。
The pathogenic power of 2.5 pathogens compares
The 36h that bacterial strain GD2 is infected after healthy leaves starts scab occur, and GD3 starts to produce scab, GD1 in 38h
Spot is then produced after 40h, it can be seen that, it is fastest that pathogen GD2 causes a disease, and is secondly bacterial strain GD2.Three kinds of pathogenic 2d of pathogens
Result afterwards as shown in Figure 11~13, a diameter of 0.24cm~0.67cm of leaf spot lesion that bacterial strain GD1 infects, generally less than GD2
The leaf spot lesion diameter 0.82cm~1.25cm for infecting, and a diameter of 0.41cm~1.19cm of leaf spot lesion that GD3 infects, explanation
The causative effect of bacterial strain GD2 is most strong, is secondly GD3.Consider with two factors of Lesion size the time required to comprehensive beginning of causing a disease, three
The pathogenic power for planting pathogen is followed successively by Alternaria tenuissima>Rod method>Phomopsis.
Embodiment 2 (cape jasmine leaf blight bacteriostatic agent screening)
1. materials and methods
1.1 strains testeds
Cape jasmine leaf blight pathogen is that embodiment 1 is separately cultured by conventional organization partition method in pathological tissues and obtains, and is purified
It is standby afterwards.Respectively Phomopsis (Phomopsis sp.), (Alternaria tenuissima Alternaria tenuissima (Fr.)
Wiltshire), rod method (Alternaria alternata (Fries) Keissler).
1.2 reagent agents
Reagent agent is commercially available, is shown in Table 1.
The reagent agent of table 1
The indoor Study on Fungicide Toxicity of 1.3 pathogens
Antibacterial medicament is determined to three kinds of inhibitory action of pathogen mycelial growth by mycelial growth rate method.Weigh certain
The reagent agent of quality, the mother liquor of certain concentration is configured to sterilized distilled water, is diluted by required concentration when using,
1mL liquids are drawn according to multiple dilutions method liquid-transfering gun to be added in the aseptic PDA culture mediums of 100mL, make reagent agent in training
Required concentration is reached in foster base.Each pathogen 6mm mycelia block is inoculated into the PDA cultures added with the bacteriostatic of required concentration respectively
In base, plus 1mL sterilized waters, in culture medium, same method inoculation is control group, and each treatment sets 3 repetitions, operates in nothing
Carried out under the conditions of bacterium.Constant temperature dark culturing in 30 DEG C of incubators is placed in, colony diameter is measured using crossing method after 5d, calculated
Bactericide sets up virulence regression equation to the inhibiting rate of mycelial growth, calculates EC50, and relatively more each reagent agent is to pathogen
Rejection ability.
Bacteriostasis rate=(control bacterium colony net diameter-treatment bacterium colony net diameter)/(control bacterium colony net diameter -0.6) 100%.
The logarithm value of drug concentration is X, and the probit value of mycelial growth relative inhibition is Y, and toxicity regression is calculated with spss19.0 softwares
Equation, coefficient correlation (r) and EC50。
2 results and analysis
Inhibition of the 2.1 different bactericide to Phomopsis
By 15 kinds of inhibitions that be can be seen that to the Toxicity Determination of Phomopsis for examination bactericide between different agents
Significant difference, is shown in Table 2.Optimal antibacterial medicament is carbendazim, EC50It is 0.19 μ gmL-1, secondly it is five nitre carbendazim, methyl sulphur
Bacterium spirit, thiophanate methyl, Bravo, EC50Respectively 0.35 μ gmL-1、2.15μg·mL-1、2.34μg·mL-1、3.04μg·
mL-1.Streptomysin, terramycin, oligosaccharides catenin fungistatic effect it is worst, EC50In 1000 μ gmL-1More than, it is not suitable for big
The disease control of area.In this experiment, the growth to Phomopsis is respectively provided with different degrees of suppression work to remaining reagent agent
With the coefficient correlation of virulence regression equation is presented the level of signifiance, and equation of linear regression is set up.15 kinds of reagents are to Phomopsis
Virulence size sequence is：Carbendazim>Five nitre carbendazim>Thiophanate-methyl>Thiophanate methyl>Bravo>Hexaconazole>Sulfuric acid celebrating is big
Mycin>Thiram>Tricyclazole>Pungent bacterium moroxydine>Zineb>Streptomycin sulphate>Oligosaccharides catenin>Terramycin>Streptomysin.This class
Topic research has only carried out the indoor measurement of pathogen preventive effect to bactericide, and crop field preventive effect need further experimental study.
2 15 kinds of bactericide of table compare the virulence of Phomopsis
Inhibition of the 2.2 different bactericide to Alternaria tenuissima
15 kinds of bactericide for trying can be seen that the suppression effect of different agents to the Toxicity Determination result of Alternaria tenuissima
Substantially, concrete outcome is shown in Table 3 to fruit difference.Wherein with the fungistatic effect of hexaconazole preferably, EC50It is 9.33 μ gmL-1, secondly it is
Thiophanate methyl, Bravo, thiram, zineb, EC50Respectively 56.89 μ gmL-1、74.29μg·mL-1、77.27μg·
mL-1、114.55μg·mL-1.Streptomysin, oligosaccharides catenin, streptomycin sulphate fungistatic effect it is worst, EC50Higher than 1000 μ g
mL-1, practical value is small.Under this various concentrations tested, the growth of pathogen is subject to different degrees of to remaining reagent agent
Inhibitory action, the coefficient correlation of virulence regression equation is presented the level of signifiance.Virulence size of 15 kinds of reagents to Alternaria tenuissima
Order is：Hexaconazole>Thiophanate methyl>Bravo>Thiram>Zineb>Tricyclazole>Pungent bacterium moroxydine>Terramycin>Carbendazim
>Five nitre carbendazim>Gentamicin sulphate>Thiophanate-methyl>Streptomycin sulphate>Oligosaccharides catenin>Streptomysin.
3 15 kinds of bactericide of table compare the virulence of Alternaria tenuissima
Inhibition of the 2.3 different bactericide to rod method
The inhibition of different agents is poor to be can be seen that to the indoor virulence of rod method by the 15 kinds of bactericide determined for examination
It is different obvious, it is shown in Table 4.Wherein with the fungistatic effect of hexaconazole preferably, EC50It is 14.45 μ gmL-1, secondly it is Bravo, methyl
Thiophanate, thiram, zineb, EC50Respectively 40.92 μ gmL-1、47.53μg·mL-1、67.92μg·mL-1、73.52μ
g·mL-1.Streptomysin, oligosaccharides catenin fungistatic effect it is worst.Remaining reagent agent under various concentrations, the life to pathogen
Length plays different degrees of inhibitory action, and the coefficient correlation of virulence regression equation is presented the level of signifiance, i.e. linear regression side
Cheng Chengli.15 kinds of reagents are to the virulence size sequence of rod method：Hexaconazole>Bravo>Thiophanate methyl>Thiram>Dai Sen
Zinc>Tricyclazole>Streptomycin sulphate>Pungent bacterium moroxydine>Terramycin>Gentamicin sulphate>Carbendazim>Five nitre carbendazim>Methyl sulphur
Bacterium spirit>Oligosaccharides catenin>Streptomysin.
4 15 kinds of bactericide of table compare the virulence of rod method
Control of plant disease depends on chemical control.When medicament is selected, it is to examine that the pathogen of medicament kills ability
The principal element of worry, and the Toxicity Determination result of bactericide can clearly illustrate that effect of the bactericide to pathogen is big
It is small, the influence of the aspect such as medicament economy, safe efficient should be considered during the use of agricultural chemicals, agricultural chemicals is played maximum effect
With, and unfavorable factor is preferably minimized.This result of the test reflects 15 kinds of different type bactericide to three kinds of suppression of pathogen
Effect.
15 kinds of bactericide have larger difference to the inhibitory action of each pathogen, different agents to the inhibitory action of pathogen not
Together, also result in the difference of valid density.Effective dose 50 is the most important index for judging medicament to pathogen inhibitory action.Together
Plant bactericide and very big difference is there is also to the fungistatic effect of three kinds of cape jasmine leaf blight pathogens, the optimal of such as Phomopsis supplies examination
Medicament is carbendazim, EC50It is 0.19 μ gmL-1, and carbendazim is to the EC of Alternaria tenuissima50Reach 396.28 μ gmL-1, it is right
Rod method then reaches 481.95 μ gmL-1, carbendazim is thousands of times of other two kinds of pathogens to the effect of Phomopsis.Soil is mould
The biological agents such as element, streptomysin, oligosaccharides catenin are worst to the inhibition of Phomopsis.Alternaria tenuissima and rod method are most
Good reagent agent is hexaconazole, contrasts toxicity test result of each medicament to both pathogens, it can be seen that medicament of the same race
Similar to the field efficacy of the two, reason is probably because the two is equal category bacterial strain, the mechanism of action and suppression of the medicament to the two
Process processed is identical.Streptomysin, oligosaccharides catenin are worst to the inhibition of Alternaria tenuissima and rod method.Compare 5 kinds of biological medicaments
Agent and 10 kinds of chemical agents find that the preventive effect of biological agent is generally anti-less than chemical agent to three kinds of fungistatic effects of pathogen
Effect.Biological pesticide is mainly characterized in that environmental protection, low toxicity, and residues of pesticides rate is low, comparatively the effect of chemical pesticide more preferably, but not
Beneficial to environmental protection, residues of pesticides rate is high, so needing to consider many aspects in the selection of applying pesticides.
Embodiments of the invention have been described in detail above, but the content is only presently preferred embodiments of the present invention,
It is not intended to limit the invention.All any modification, equivalent and improvement made in application range of the invention etc., all should
It is included within protection scope of the present invention.
Claims (10)
1. a kind of prevention and controls for cape jasmine leaf blight disease, it is characterised in that Phomopsis are applied to cape jasmine plant antibacterial
Agent, Alternaria tenuissima bacteriostatic agent, alternaric bacteria bacteriostatic agent.
2. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 1, it is characterised in that the plan stem
It is N- (2- benzimidazoles base)-methyl carbamate to select mould bacteriostatic agent.
3. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 2, it is characterised in that N- (2- benzene a pair of horses going side by sides
Imidazole radicals)-methyl carbamate concentration be 0.19 μ g/mL.
4. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 1, it is characterised in that the thin pole
Rod method bacteriostatic agent is hexaconazole.
5. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 4, it is characterised in that hexaconazole
Concentration is 9.33 μ g/mL.
6. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 1, it is characterised in that the chain lattice
Spore bacterium bacteriostatic agent is hexaconazole.
7. a kind of prevention and controls for cape jasmine leaf blight disease according to claim 6, it is characterised in that hexaconazole
Concentration is 14.45 μ g/mL.
8. a kind of composition for claim 1 methods described, it is characterised in that said composition includes N- (2- benzimidazoles
Base)-methyl carbamate and hexaconazole.
9. composition according to claim 8, it is characterised in that wherein N- (2- benzimidazoles base)-methyl carbamate
Concentration is 15 μ gmL-1。
10. composition according to claim 8, it is characterised in that wherein the concentration of hexaconazole is 500 μ gmL-1。
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