CN117581868A - Bactericide for preventing and treating kiwi fruit canker and citrus canker and application - Google Patents
Bactericide for preventing and treating kiwi fruit canker and citrus canker and application Download PDFInfo
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- CN117581868A CN117581868A CN202311582049.8A CN202311582049A CN117581868A CN 117581868 A CN117581868 A CN 117581868A CN 202311582049 A CN202311582049 A CN 202311582049A CN 117581868 A CN117581868 A CN 117581868A
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- Prior art keywords
- bactericide
- active ingredient
- canker
- kiwi fruit
- agent
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Classifications
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a bactericide for preventing and treating kiwi fruit canker and citrus canker and application thereof. The invention can radically and effectively inhibit citrus canker, has quick response and no residue, improves the quality of citrus, and has better effect compared with 77% basic copper sulfate water dispersible granules. The invention can radically and effectively inhibit kiwi fruit canker, has quick response and no residue, improves kiwi fruit quality, and has better effect compared with 5% kasugamycin aqua.
Description
Technical Field
The invention relates to the field of botanical bactericides, in particular to a bactericide for preventing and treating kiwi fruit canker and citrus canker and application thereof.
Background
The damage of plant pathogenic bacteria seriously threatens the yield and quality of environmental crops, wherein bacterial diseases such as kiwi canker, citrus yellow dragon disease, tomato bacterial wilt, rice bacterial leaf blight and the like have serious influence on industry, not only lead to yield reduction and harvest loss, but also possibly destroy gardens and realize harvest failure. At present, the main bacterial diseases prevention and control methods include inorganic copper, organic copper, antibiotics, microbial pesticides and the like. Copper preparations widely applied to bacterial diseases in production comprise copper hydroxide, copper regen, basic copper sulfate and the like, the active ingredients have single action targets, and the bacteria have high drug resistance after long-term use. In addition, the inorganic copper preparation is easy to generate phytotoxicity at high temperature, and stimulates the occurrence of red spiders; bacterial diseases show interactive resistance to organic copper preparations such as oxine and the like and inorganic copper preparations, and the field control effect is poor; the microbial pesticide such as bacillus subtilis and the like has extremely unstable control effect and poor quick-acting property; the antibiotic medicaments such as tetramycin, kasugamycin, zhongshengmycin and the like have weak rain wash resistance and poor permeability, do not have outstanding protection effect on crops in the condition of continuous raininess in the south, and cannot be mixed with alkaline pesticides. The dosage forms registered by the existing products are mainly wettable powder, emulsifiable concentrate, water aqua, soluble liquid, water dispersible granules and the like, the dosage forms can not conduct active ingredients to bast layers, the permeability and the rain wash resistance are not ideal, and the control effect on bacterial diseases is not good. Therefore, the development of the bactericide with unique action mechanism and excellent biological activity is always a research hot spot in the field of new pesticides, and particularly has great prospect in the aspects of excavating plant source active ingredients with high antibacterial activity and developing environment-friendly agricultural bactericides.
Oregano oil is an active essential oil component extracted from Oregano plant material, and has various biological functions including sterilization, deinsectization, disinfection, antioxidation, corrosion prevention, etc. The oregano oil is a safe, efficient, green and pure natural high-activity ingredient traditional Chinese medicine additive without incompatibility, is one of feed medicine additives approved for use in China, and comprehensively evaluates the safety of animals and non-target organisms. The Chinese patent application CN112219865A discloses that the composition has stronger inhibition effects on corn leaf spot mold, rhizoctonia solani, sclerotinia sclerotiorum, botrytis cinerea, wheat gibberella, gummi citrulli, cotton fusarium wilt, rice blast germ, phytophthora capsici, apple anthracnose germ, fusarium oxysporum, oat fusarium, horsetail fusarium, colletotrichum oxysporum, fusarium solani, alternaria tenuis, citrus canker germ and potato black shank germ, and particularly has excellent inhibition effects on the inhibition of rhizoctonia solani. However, the disclosed patent uses the biological activity measurement result of oregano oil dispersion system on bacterial disease citrus canker pathogen and potato black shank pathogen, the inhibition rate is 100% at the concentration of 250 mug/ml, and the inhibition rate is only 63.2% and 39.9% at the concentration of 125 mug/ml. However, no report on the use of oregano oil for preventing and treating kiwi fruit canker is seen.
Carvacrol is the main component of oregano oil, can be separated and purified from the oregano oil, and can also be extracted from various aromatic plant raw materials such as oregano, thyme, and guava. As the carvacrol has no toxic or side effect, no residue and no pollution to the environment, the common-effect spice is added into cosmetics and also used as a bacteriostatic agent and a preservative for food preservation, and medical research discovers that carvacrol also has biological activities of inhibiting tumor, resisting inflammation, resisting oxidation, regulating blood sugar, resisting parasites and the like. The Chinese pesticide information net bulletin, carvacrol (5% carvacrol aqua, accession number PD 20171458) is registered for diaphorina citri, citrus red spider, rice planthopper, tobacco virus disease, tobacco brown spot, tobacco powdery mildew, tomato gray mold, tea leafhopper, potato late blight, cucumber bacterial angular leaf spot and the like. The Chinese patent application CN109006855A discloses an insecticidal composition containing sulfoxaflor and carvacrol, wherein the active ingredients consist of sulfoxaflor and carvacrol, and the weight ratio of the sulfoxaflor to the carvacrol is 1-40:1. The pesticide composition is suitable for controlling tea leafhoppers, can delay the development of pest resistance, reduce the application times, improve the control effect and simultaneously is beneficial to reducing chemical pesticide residues. The Chinese patent application CN108605939A discloses a bactericidal composition containing carvacrol and ethirimol, wherein the active ingredients consist of carvacrol and ethirimol in a weight ratio of 1:10-30. The bactericidal composition reasonably mixed by carvacrol and ethirimol is used for preventing and treating powdery mildew of crops, has obvious synergistic effect and long lasting period, and is beneficial to prolonging the life cycle of ethirimol. The Chinese patent application CN 109090110A also discloses a sterilization composition containing carvacrol and flufenpyroximate as active ingredients, which comprises common auxiliary agents, carvacrol and flufenpyroximate, wherein the mass ratio of the carvacrol to the flufenpyroximate is 3:1-1:5, and the preferential ratio is 1:1-1:3. After being compounded with the fenpyroximate according to a certain proportion, the carvacrol and the fenpyroximate have synergistic effect on pathogenic fungi such as tomato leaf mold, and are suitable for preventing and treating crop diseases caused by the pathogenic fungi such as tomato leaf mold. However, no report on controlling citrus canker and kiwi canker diseases by carvacrol exists so far.
The invention of China patent CN108835110B discloses a pesticide slow release microsphere and a preparation method thereof, the pesticide slow release microsphere is composed of a core material and a wall material, wherein the mass ratio of the core material to the wall material is 1 (1-6), the core material is a pesticide raw material, and the wall material is composed of polylactic acid (PLA) and polybutylene succinate (PBS) (the mass ratio is 1:1-9).
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides an agricultural bactericide and application thereof, which can be used for preventing and treating bacterial pathogen harm, in particular to a bactericide for preventing and treating kiwi fruit canker and citrus canker and application thereof, has quick response, no residue and improves the quality of kiwi fruits and citrus.
In order to solve the technical problems, the invention adopts the following technical scheme: a bactericide for preventing and treating kiwi canker and citrus canker, which comprises an active ingredient, wherein the active ingredient is one or more of oregano oil and carvacrol.
The invention provides one or more of origanum oil and carvacrol which are plant extracts for inhibiting pathogenic bacteria of citrus canker and kiwi fruit canker, and the occurrence of the citrus canker and kiwi fruit canker is controlled by inhibiting the growth and propagation of the pathogenic bacteria of the citrus canker and kiwi fruit canker.
Compared with other purely natural plant source biological preparation pesticides or chemical pesticides, the oregano oil and carvacrol in the application have excellent control effects on citrus canker and kiwi fruit canker.
In a preferred embodiment of the present invention, the bactericide further comprises an auxiliary agent, wherein the auxiliary agent comprises one or more of a surfactant, a filler, a solvent, a carrier, an emulsifier, a dispersing agent, a foaming agent, a defoaming agent, a wetting agent, a thickening agent, a penetrating agent, a stabilizer, an antifreezing agent, a film forming agent, a color paste and an essence, and the active ingredient accounts for 0.5-95.0% of the total mass of the bactericide, preferably, the active ingredient accounts for 1.0-50.0% of the total mass of the bactericide, and further preferably, the active ingredient accounts for 5.0-25.0% of the total mass of the bactericide.
According to embodiments of the present invention, the germicides may also include stabilizers such as low temperature stabilizers, preservatives, antioxidants, light stabilizers or other aids that enhance chemical and/or physical stability.
According to embodiments of the present invention, the germicides may be prepared by known means, such as by mixing oregano oil or carvacrol with adjuvants.
According to an embodiment of the present invention, the active ingredient is mixed with an adjuvant to prepare a formulation suitable for agricultural use, which formulation may be selected from one or more of an aqueous emulsion, a microemulsion, an oil emulsion, an emulsifiable concentrate, a latex, a paste, a dispersible agent, a soluble agent, an oil agent, a granule, a wettable powder, a water dispersible granule, a suspension agent, a microcapsule suspension agent, an oil suspension agent, a dispersible oil suspension agent, a seed treatment suspension agent.
The germicides of the present invention may be used to treat plants or plant parts. Or allowing the active ingredient to act on its environment, habitat or storage space, for example by dipping, spraying, atomizing, drip irrigation, injection, flushing, evaporation, spreading, painting, coating (for example by application to seeds) or dressing.
In some embodiments of the invention, the dosage form of the germicide is a microcapsule suspension. The bactericide comprises a core material and a wall material in a mass ratio of 1:5-5:1, wherein the core material is an active ingredient, and the wall material comprises polybutylene succinate and polyhydroxybutyrate in a mass ratio of 1:4-4:1; the active ingredient is wrapped in the wall material, preferably, the bactericide comprises a core material and a wall material with the mass ratio of 1:1-3:1, and the mass ratio of the wall material consisting of polybutylene succinate and polyhydroxybutyrate is 1:1-1:1.5.
The active ingredients of oregano oil and carvacrol in the invention belong to plant extracts, are extremely easy to oxidize and volatilize under natural conditions, and can be directly applied and exposed in the air in field application, so that the amount of active ingredients acting on a target is small due to degradation or escape of the active ingredients, and the biological activity is reduced. The microcapsule suspending agent prepared from the active ingredients can wrap the active ingredients, avoid volatilization and degradation caused by ultraviolet rays, improve the bioavailability and obviously improve the field prevention effect. The choice of microcapsule wall material has a great relation with the active ingredient to be coated, and influences whether the active ingredient can be fully coated, the proportion in the patent is the optimal proportion of origanum oil and carvacrol relative to the wall material, and too high or too low can lead to insufficient coating of the active ingredient.
Aiming at the core material of the application, the wall material prepared from polylactic acid (PLA) and polybutylene succinate (PBS) in the patent of the invention with the publication number of CN108835110B is easy to form capsules, but the prepared microcapsule is irregular in appearance, easy to generate holes, low in encapsulation efficiency, quick in active ingredient release and short in duration. Compared with the wall material or other wall materials in the patent with the publication number of CN108835110B, the microcapsule prepared from the Polyhydroxybutyrate (PHB) and the polybutylene succinate (PBS) has good encapsulation condition, regular spherical shape, regular shape, no agglomeration phenomenon, higher encapsulation efficiency and drug loading rate and better drug effect.
The preparation method of the bactericide comprises the following steps:
(1) Preparation of an oil phase: dissolving the active ingredients, polyhydroxybutyrate and polybutylene succinate in a low-boiling-point organic solvent to obtain an oil phase containing the active ingredients and a carrier;
(2) Preparation of an aqueous phase: completely dissolving an emulsifying dispersant in water to obtain a water phase;
(3) Preparation of microcapsules: adding the oil phase containing the active ingredients and the carrier in the step (1) into the water phase in the step (2), shearing at room temperature at high speed, obtaining microemulsion after shearing is finished, stirring the microemulsion at a set temperature until the organic solvent is completely volatilized, and obtaining the drug-loaded microcapsule after centrifugation and drying;
(4) Microcapsule suspending agent preparation: measuring the drug loading quantity of the active ingredients, and then measuring the mass of the microcapsule, the wetting agent, the dispersing agent, the thickening agent, the permeation enhancer and the stabilizing agent according to the proportion of the microcapsule suspending agent, and mixing the materials in a container to prepare the preparation through shearing and dispersing.
The invention also discloses application of the bactericide in preventing and treating kiwi fruit canker diseases.
In a preferred embodiment of the invention, the active ingredient of the bactericide is carvacrol, the mass concentration of the active ingredient in the bactericide is 4-20 mg/L, and the EC of the active ingredient in the bactericide 50 10.55mg/L.
In a preferred embodiment of the invention, the active ingredient of the bactericide is oregano oil, the mass concentration of the active ingredient in the bactericide is 5-40 mg/L, and the EC of the active ingredient in the bactericide 50 15.38mg/L.
EC of active ingredient in the bactericide 50 The values are determined by turbidimetry under laboratory conditions.
In a preferred embodiment of the invention, the active ingredient of the bactericide is carvacrol, and the mass concentration of the bactericide applied in the field is 300-800 mg/L, preferably the mass concentration of the bactericide applied in the field is 500mg/L.
In a preferred embodiment of the invention, the active ingredient of the bactericide is oregano oil, the mass concentration of the bactericide applied in the field is 300-800 mg/L, and preferably the mass concentration of the bactericide applied in the field is 500mg/L.
The preferred mass concentration of the bactericide is obtained by using the bactericide in field test measurement.
The mass concentration range of the active ingredients in the bactericide can obviously inhibit the growth and reproduction of kiwi fruit canker pathogens.
The invention also discloses application of the bactericide in preventing and treating citrus canker.
In a preferred embodiment of the invention, the active ingredient of the bactericide is carvacrol, the mass concentration of the active ingredient in the bactericide is 3-15 mg/L, and the EC of the active ingredient in the bactericide 50 7.52mg/L.
In a preferred embodiment of the invention, the active ingredient of the bactericide is carvacrol, the mass concentration of the bactericide is 300-800 mg/L, and preferably, the mass concentration of the bactericide is 500mg/L.
The preferred mass concentration of the bactericide is obtained by using the bactericide in field test measurement.
The mass concentration range of the active ingredients in the bactericide can obviously inhibit the growth and reproduction of citrus canker pathogens.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention can effectively inhibit the citrus canker, has quick response and no residue, improves the quality of citrus, and has better effect compared with 77% basic copper sulfate water dispersible granules.
(2) The invention can effectively inhibit kiwi fruit canker, has quick response and no residue, improves kiwi fruit quality, and has better effect compared with 5% kasugamycin aqua.
Drawings
FIG. 1 is a graph showing the results of measuring the indoor toxicity of carvacrol against kiwi fruit canker pathogens by turbidimetry in example 1; the color of the bacterial liquid is gradually clear along with the increase of the concentration of the active ingredients, which shows that the carvacrol with higher concentration can obviously inhibit the growth and reproduction of kiwi fruit canker.
FIG. 2 is a photograph of 10% carvacrol microcapsule suspension prepared in example 2 after administration for preventing and treating kiwi fruit canker; the plants after the application have no obvious canker hazard symptoms, which shows that the 10% carvacrol microcapsule suspending agent has obvious field control effect.
FIG. 3 is a graph comparing the post-drug administration of 4% kasugamycin aqueous solution for preventing and treating kiwi fruit canker; the blades have obvious canker hazard symptoms after the kasugamycin is taken, which shows that the 4% kasugamycin aqua has weaker capability of preventing and treating kiwi fruit canker.
FIG. 4 is a graph showing the results of nephelometric determination of carvacrol against citrus canker pathogens in example 3; the color of the bacterial liquid is gradually clear along with the increase of the concentration of the active ingredients, which shows that the carvacrol with higher concentration can obviously inhibit the growth and reproduction of kiwi fruit canker.
FIG. 5 is a photograph of 10% carvacrol microcapsule suspension prepared in example 4 after administration for the prevention and treatment of citrus canker; the plants after the application have no obvious canker hazard symptoms, which shows that the 10% carvacrol microcapsule suspending agent has obvious field control effect.
FIG. 6 is a photograph of a comparative 77% basic copper sulfate water dispersible granule used for controlling citrus canker; obvious ulcer disease hazard symptoms exist on leaves after the 77% basic copper sulfate water dispersible granule is used, which shows that the 77% basic copper sulfate water dispersible granule has weaker capability of preventing and treating kiwi fruit ulcer disease.
FIG. 7 is a graph showing the results of measuring indoor toxicity of oregano oil against kiwi fruit canker by turbidimetry in example 5; the color of the bacterial liquid is gradually clear along with the increase of the concentration of the active ingredients, which shows that the oregano oil with higher concentration can obviously inhibit the growth of kiwi fruit canker.
FIG. 8 is a photograph of 10% oregano oil microcapsule suspension prepared in example 6 after use for controlling kiwi fruit canker; the plants after the pesticide are not provided with obvious canker hazard symptoms, which shows that the 10% oregano oil microcapsule suspending agent has obvious field control effect.
FIG. 9 is a graph comparing the post-drug administration of 4% kasugamycin aqueous solution for preventing and treating kiwi fruit canker; the blades have obvious canker hazard symptoms after the kasugamycin is taken, which shows that the 4% kasugamycin aqua has weaker capability of preventing and treating kiwi fruit canker.
Detailed Description
Example 1:
method for testing the in vitro inhibitory activity of carvacrol against the kiwi ulcer pathogen pseudomonas syringae kiwi fruit pathogenic variant Pseudomonas syringae pv.actinidia e (Psa):
biological test material: the kiwi fruit canker pathogen is a physiological micro-species strain, namely the yellow kiwi fruit subspecies of kiwi fruit, which is collected, separated and identified by the inventor. The strain is activated on LB solid medium and cultured for 3 days at 28 ℃ for later use.
Experimental instrument and consumable: ultraviolet spectrometer, constant temperature incubator, constant temperature shaking table, centrifuging tube, rifle head.
A step of biological measurement: preparing a culture medium: LB liquid medium: 10g of tryptone, 5g of yeast extract, 10g of sodium chloride, and adjusting the pH to 7.4 by using distilled water to 1000 ml.
Preparation of bacterial suspension: activating the strain on an LB solid medium, culturing for 3 days at 28 ℃ in an inverted mode, slightly scraping a small amount of bacterial colonies on an ultra-clean workbench by using a gun head pipe, culturing on a shaking table at 28 ℃ under 200rpm until the bacterial colonies are completely dispersed and have no massive precipitate, and diluting kiwi fruit canker bacteria into a bacterial suspension with a light absorption value of 0.1 at 600nm of an ultraviolet spectrophotometer by using sterile water.
And determining the toxicity of the active ingredients to kiwi fruit canker by using a turbidimetry method. Setting 5 concentrations of active ingredientAnd (3) setting 100mLLB culture solutions for each gradient, adding 100 mu L of prepared kiwi fruit canker fungus liquid, placing into a constant temperature oscillator, culturing for 24 hours at 28 ℃ and 180r/min, taking LB culture medium without active ingredients as CK, and repeating for 3 times for each test group. Then measuring the solubility of the bacterial liquid at 600nm by using an ultraviolet light spectrometer, and calculating a virulence regression equation and EC by using a DPS data processing system 50 Values and correlation coefficients. The growth inhibition rate was calculated according to formula 1-1:
table 1 determination of indoor toxicity results of carvacrol on kiwi fruit canker by turbidimetry
Example 2:
a preparation for preventing and treating kiwi fruit canker is provided. The adhesive is prepared from the following components in parts by weight: 98 parts of deionized water, 2 parts of polyvinyl alcohol, 10 parts of carvacrol, 2.25 parts of polybutylene succinate (PBS), 2.75 parts of Polyhydroxybutyrate (PHB) and 30 parts of methylene dichloride. During preparation, 98g of deionized water is measured in an conical flask, 2g of polyvinyl alcohol is slowly added, the mixture is heated to 60 ℃ by a digital display magnetic stirrer and stirred at a constant temperature and high speed to be completely dissolved, and the mixture is cooled to room temperature to prepare a water phase. With PBS: PHB (9:11) is used as a wall material, carvacrol serving as an active ingredient is used as a core material, and the core-wall ratio is 2:1. 2.25g of polybutylene succinate (PBS) and 2.75g of Polyhydroxybutyrate (PHB) are dissolved in 30mL of dichloromethane, and the mixture is fully stirred on a magnetic stirrer, and then carvacrol serving as an active ingredient is added into the mixture to be continuously stirred and dissolved, so that an oil phase can be obtained. After the oil phase is poured into the water phase, a T25 digital display disperser is adopted to emulsify and shear for 5min at the speed of 8000r/min, so that stable oil-in-water mixed emulsion can be prepared, and the oil-in-water mixed emulsion is stirred for 6-7 h at the speed of 600r/min on a magnetic stirrer until the solvent dichloromethane is completely volatilized, and the microcapsule is solidified. And separating the microcapsules, and measuring the drug loading rate of the active ingredients. The microcapsule, wetting agent, dispersing agent, thickening agent, permeation enhancer and stabilizing agent are metered according to the proportion of the microcapsule suspending agent, and are combined in a container to be sheared and dispersed to prepare the preparation. When in use, the preparation and water are mixed according to the mass ratio of 1:200, and spraying kiwi fruit plants.
The preparation for preventing and treating kiwi fruit canker prepared in the above example 2 was subjected to a control experiment according to the use method. The results are shown in table 2 below:
table 2 results of field efficacy investigation of 10% carvacrol microcapsule suspension on kiwi canker
Example 3:
the method for testing the in-vitro inhibition activity of carvacrol on citrus canker pathogenic bacteria comprises the following steps:
biological test material: the invention relates to a citrus canker pathogen strong pathogenicity physiological micro-species, namely, xanthomonas citri subspecies (Xanthomonas citri subsp. Citri, xcc) LYGJ3-5, which is collected, separated and identified by the inventor. The strain is activated on LB solid medium and cultured for 3 days at 28 ℃ for later use.
Experimental instrument and consumable: ultraviolet spectrometer, constant temperature incubator, constant temperature shaking table, centrifuging tube, rifle head.
A step of biological measurement: preparing a culture medium: LB liquid medium: 10g of tryptone, 5g of yeast extract, 10g of sodium chloride, and adjusting the pH to 7.4 by using distilled water to 1000 ml.
Preparation of bacterial suspension: activating the strain on an LB solid medium, culturing for 3 days at 28 ℃ in an inverted mode, slightly scraping a small amount of bacterial colonies on an ultra-clean workbench by using a gun head pipe, culturing on a shaking table at 28 ℃ under the vibration of 200rpm until the bacterial colonies are completely dispersed and have no massive precipitate, and diluting citrus canker bacteria into bacterial suspension with a light absorption value of 0.1 at 600nm of an ultraviolet spectrophotometer by using sterile water.
The toxicity of the active ingredient to the citrus canker is determined by turbidimetry. 5 concentration gradients are set for the active ingredients, 100mL of LB culture solution is set for each gradient, 100 mu L of prepared citrus canker fungus solution is added, the mixture is placed into a constant temperature oscillator for culturing for 24 hours at 28 ℃ and 180r/min, LB culture medium without the active ingredients is used as CK, and 3 repetitions are carried out for each test group. Then measuring the solubility of the bacterial liquid at 600nm by using an ultraviolet light spectrometer, and calculating a virulence regression equation and EC by using a DPS data processing system 50 Values and correlation coefficients. Growth inhibition was calculated according to formula 3-1:
table 3 determination of indoor virulence results of carvacrol on citrus canker by turbidimetry
Example 4:
a preparation for preventing and treating citrus canker is provided. The adhesive is prepared from the following components in parts by weight: 98 parts of deionized water, 2 parts of polyvinyl alcohol, 10 parts of carvacrol, 2.25 parts of polybutylene succinate (PBS), 2.75 parts of Polyhydroxybutyrate (PHB) and 30 parts of methylene dichloride. During preparation, 98g of deionized water is measured in an conical flask, 2g of polyvinyl alcohol is slowly added, the mixture is heated to 60 ℃ by a digital display magnetic stirrer and stirred at a constant temperature and high speed to be completely dissolved, and the mixture is cooled to room temperature to prepare a water phase. With PBS: PHB (9:11) is used as a wall material, carvacrol serving as an active ingredient is used as a core material, and the core-wall ratio is 2:1. 2.25g of polybutylene succinate (PBS) and 2.75g of Polyhydroxybutyrate (PHB) are dissolved in 30mL of dichloromethane, and the mixture is fully stirred on a magnetic stirrer, and then carvacrol serving as an active ingredient is added into the mixture to be continuously stirred and dissolved, so that an oil phase can be obtained. After the oil phase is poured into the water phase, a T25 digital display disperser is adopted to emulsify and shear for 5min at the speed of 8000r/min, so that stable oil-in-water mixed emulsion can be prepared, and the oil-in-water mixed emulsion is stirred for 6-7 h at the speed of 600r/min on a magnetic stirrer until the solvent dichloromethane is completely volatilized, and the microcapsule is solidified. And separating the microcapsules, and measuring the drug loading rate of the active ingredients. The microcapsule, wetting agent, dispersing agent, thickening agent, permeation enhancer and stabilizing agent are metered according to the proportion of the microcapsule suspending agent, and are combined in a container to be sheared and dispersed to prepare the preparation. When in use, the preparation and water are mixed according to the mass ratio of 1:200, and spraying kiwi fruit plants.
The bactericide for preventing and treating citrus canker prepared by the preparation method of the above example 4 was subjected to a control experiment on a navel orange planting base according to the use method. As shown in table 4 below:
table 4 results of field efficacy investigation of 10% carvacrol microcapsule suspension on citrus canker
Example 5:
the method for testing the in-vitro inhibition activity of oregano oil on the actinidia canker pathogen pseudomonas syringae and actinidia chinensis pathogenic variant Pseudomonas syringae pv.
Biological test material: the kiwi fruit canker pathogen is a physiological micro-species strain, namely the yellow kiwi fruit subspecies of kiwi fruit, which is collected, separated and identified by the inventor. The strain is activated on LB solid medium and cultured for 3 days at 28 ℃ for later use.
Experimental instrument and consumable: ultraviolet spectrometer, constant temperature incubator, constant temperature shaking table, centrifuging tube, rifle head.
A step of biological measurement: preparing a culture medium: LB liquid medium: 10g of tryptone, 5g of yeast extract, 10g of sodium chloride, and adjusting the pH to 7.4 by using distilled water to 1000 mL.
Preparation of bacterial suspension: activating the strain on an LB solid medium, culturing for 3 days at 28 ℃ in an inverted mode, slightly scraping a small amount of bacterial colonies on an ultra-clean workbench by using a gun head pipe, culturing on a shaking table at 28 ℃ under 200rpm until the bacterial colonies are completely dispersed and have no massive precipitate, and diluting kiwi fruit canker bacteria into bacterial suspension with a light absorption value of 0.1 at 600nm of an ultraviolet spectrophotometer by using sterile water.
And determining the toxicity of the active ingredients to kiwi fruit canker by using a turbidimetry method. Setting 5 concentration gradients of active ingredients, setting 100mL of LB culture solution for each gradient, adding 100 mu L of prepared kiwifruit canker fungus liquid, placing into a constant-temperature oscillator, culturing for 24h at 28 ℃ and 180r/min, taking LB culture medium without active ingredients as CK, and repeating for 3 times for each test group. Then measuring the solubility of the bacterial liquid at 600nm by using an ultraviolet light spectrometer, and calculating a virulence regression equation and EC by using a DPS data processing system 50 Values and correlation coefficients. Growth inhibition was calculated according to formula 5-1:
table 5 determination of indoor virulence results of oregano oil on kiwi fruit ulcer disease by nephelometry
Example 6:
a preparation for preventing and treating kiwi fruit canker is provided. The adhesive is prepared from the following components in parts by weight: 98 parts of deionized water, 2 parts of polyvinyl alcohol, 10 parts of origanum oil, 2 parts of polybutylene succinate (PBS), 3 parts of Polyhydroxybutyrate (PHB) and 30 parts of methylene dichloride. During preparation, 98g of deionized water is measured in an conical flask, 2g of polyvinyl alcohol is slowly added, the mixture is heated to 60 ℃ by a digital display magnetic stirrer and stirred at a constant temperature and high speed to be completely dissolved, and the mixture is cooled to room temperature to prepare a water phase. With PBS: PHB (2:3) is used as a wall material, carvone serving as an active ingredient is used as a core material, and the core-wall ratio is 2:1. 2g of polybutylene succinate (PBS) and 3g of Polyhydroxybutyrate (PHB) are dissolved in 30mL of dichloromethane, and the mixture is fully stirred on a magnetic stirrer, and then the active ingredient carvone is added into the mixture to be continuously stirred and dissolved, so that an oil phase can be obtained. After the oil phase is poured into the water phase, a T25 digital display disperser is adopted to emulsify and shear for 5min at the speed of 8000r/min, so that stable oil-in-water mixed emulsion can be prepared, and the oil-in-water mixed emulsion is stirred for 6-7 h at the speed of 600r/min on a magnetic stirrer until the solvent dichloromethane is completely volatilized, and the microcapsule is solidified. And separating the microcapsules, and measuring the drug loading rate of the active ingredients. The microcapsule, wetting agent, dispersing agent, thickening agent, permeation enhancer and stabilizing agent are metered according to the proportion of the microcapsule suspending agent, and are combined in a container to be sheared and dispersed to prepare the preparation. When in use, the preparation and water are mixed according to the mass ratio of 1:200, and spraying kiwi fruit plants.
Table 6 results of field efficacy investigation of 10% oregano oil microcapsule suspension on kiwi fruit canker
Claims (10)
1. The bactericide for preventing and treating kiwi fruit canker diseases and citrus canker is characterized by comprising an active ingredient, wherein the active ingredient is one or more of oregano oil and carvacrol.
2. The bactericide for controlling citrus canker and kiwi fruit canker diseases according to claim 1, wherein the bactericide further comprises an auxiliary agent, the auxiliary agent comprises one or more of a surfactant, a filler, a solvent, a carrier, an emulsifier, a dispersant, a foaming agent, a defoaming agent, a wetting agent, a thickener, a penetrating agent, a stabilizer, an antifreezing agent, a film forming agent, color paste and essence; the content of the active ingredient is 0.5-95.0% of the total weight of the bactericide, preferably 1.0-50.0% of the total weight of the bactericide, and further preferably 5.0-25.0% of the total weight of the bactericide.
3. The bactericide for controlling citrus canker and kiwi fruit canker according to claims 1-2, characterized in that the active ingredient is mixed with an auxiliary agent to prepare a formulation suitable for agricultural use, wherein the formulation is selected from one or more of aqueous emulsion, microemulsion, oil emulsion, emulsifiable concentrate, latex, paste, ointment, dispersible liquid, soluble liquid, oil, granule, wettable powder, water dispersible granule, suspending agent, microcapsule suspending agent, oil suspending agent, dispersible oil suspending agent and seed treatment suspending agent; preferably, the dosage form is a microcapsule suspending agent, wherein the microcapsule consists of a core material and a wall material, the mass ratio of the core material to the wall material is 1:5-5:1, the core material is an active ingredient, and the wall material consists of polybutylene succinate and polyhydroxybutyrate, and the mass ratio of the polybutylene succinate to the polyhydroxybutyrate is 1:4-4:1; the active ingredient is wrapped in the wall material, preferably, the bactericide comprises a core material and a wall material with the mass ratio of 1:1-3:1, and the mass ratio of the wall material consisting of polybutylene succinate and polyhydroxybutyrate is 1:1-1:1.5.
4. Use of the bactericide according to any one of claims 1 to 3 for controlling kiwi fruit canker diseases.
5. The use of the bactericide for preventing and treating kiwi fruit canker diseases according to claim 4, wherein the active ingredient of the bactericide is carvacrol, the mass concentration of the active ingredient in the bactericide is 4-20 mg/L, and the EC of the active ingredient in the bactericide 50 10.55mg/L.
6. The use of the bactericide for preventing and treating kiwi fruit canker diseases according to claim 4, wherein the active ingredient of the bactericide is carvacrol, the mass concentration of the bactericide applied in fields is 300-800 mg/L, and preferably, the mass concentration of the bactericide applied in fields is 500mg/L.
7. The use of the bactericide for preventing and treating kiwi fruit canker diseases according to claim 4, wherein the active ingredient of the bactericide is origanum oil, the mass concentration of the active ingredient in the bactericide is 5-40 mg/L, and the EC of the active ingredient in the bactericide 50 15.38mg/L;
8. use of a fungicide according to any one of claims 1 to 3 for the control of citrus canker.
9. The use of the bactericide according to claim 8, wherein the active ingredient of the bactericide is carvacrol, the mass concentration of the active ingredient in the bactericide is 3-15 mg/L, and the EC of the active ingredient in the bactericide 50 7.52mg/L.
10. Use of a bactericide according to claim 8 for controlling citrus canker, characterized in that the active ingredient of the bactericide is carvacrol, the mass concentration of the bactericide applied in the field is 300-800 mg/L, preferably the mass concentration of the bactericide applied in the field is 500mg/L.
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