AU2021100132A4 - Biological pesticide preparation for controlling rice diseases and preparation method and use thereof - Google Patents
Biological pesticide preparation for controlling rice diseases and preparation method and use thereof Download PDFInfo
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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
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
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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
- A01N25/02—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 containing liquids as carriers, diluents or solvents
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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
- A01N25/30—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 characterised by the surfactants
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- A01N41/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a sulfur atom bound to a hetero atom
- A01N41/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a sulfur atom bound to a hetero atom containing a sulfur-to-oxygen double bond
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Abstract
The present disclosure provides a biological pesticide preparation for controlling rice diseases,
and a preparation method and use thereof. In a biological pesticide preparation for preventing rice
diseases and a preparation method thereof, the biological pesticide preparation is prepared into new
forms of environmentally-friendly pesticide oil in water (O/W) emulsion and microemulsion with
kasugamycin and allicin as active ingredients. In the present disclosure, the microemulsion and the
O/W emulsion have an optimized formula including a pH adjusting agent to enhance stability of
kasugamycin. Moreover, the present disclosure uses no organic solvent, and includes a synergist
during preparation which effectively improves efficacy of the product. The biological pesticide
preparation can be used to control rice diseases, and has outstanding effects on rice blast, rice bacterial
leaf blight and rice panicle blight, which broadens the application range of the product with an
excellent effect.
Description
The present disclosure relates to the technical field of pesticide preparations, and a biological pesticide preparation containing allicin and kasugamycin, and a preparation method and use thereof.
Allicin is an organic sulfur compound extracted from bulbs (garlic heads) of Allium Sativum, a plant in the Allium family. It also exists in onions and other plants in the Allium family. Its scientific name is diallyl thiosulfinate. The allicin is a broad-spectrum antibacterial drug for use in pharmaceutical production, which has many functions such as eliminating inflammation and virus, lowering blood pressure and blood lipids, protecting liver, and preventing diabetes. It can also be used in animal husbandry and aquaculture to improve production performance, reduce morbidity and mortality, and increase breeding efficiency.
The allicin can be further used in control of a wide variety of plant diseases and pests. As a main component thereof, an organic thioether compound has insecticidal, microbicidal and health care effects on crops, and can effectively improve crop yield and quality after application. The allicin can inhibit a variety of gram-negative and gram-positive bacteria, and has an excellent effect on tomato early blight, leaf mold and gray mold. It can effectively inhibit growth of pathogens of rape Sclerotinia stem rot, wheat sheath blight, pepper phytophthora blight, pepper anthracnose, and rice blast.
Kasugamycin is a systemic fungicide and bactericide with protective and therapeutic functions. It can inhibit the mycelial growth of rice blast fungus and prevent development of the disease.
The two agents have excellent control effects on rice blast, rice bacterial leaf blight and rice panicle blight (also known as rice bacterial blight). Combination of the two can effectively slow down drug resistance, improve the control effects and reduce pesticide usage. The compound medicaments can effectively increase use of botanical pesticides in agricultural production and reduce use of chemical pesticides to help addressing the existing pesticide residue and resistance problems. At present, there is no report on use of combination of the two in control of rice blast, rice bacterial leaf blight, or rice panicle blight.
Currently, Xiao Tian and Yao Tingshan from Southwest University in China have submitted a patent application entitled "A pharmaceutical composition for control of FructusA urantiidisease and
I a preparation method thereof' with a patent application number of 201610164135.0. This patent application provides a medicine combination for control of FructusAurantii disease with allicin and kasugamycin as main components. It discloses a preparation method and a product formula for a pesticide preparation which includes a composition of allicin and kasugamycin. It also discloses methods for preparing the composition in form of a soluble liquid and a soluble powder.
Through analysis of the above patent application, it is found that the application has the following defects: (1): Sodium carboxymethyl cellulose (CMC-Na) used in the formula is alkaline in an aqueous solution while the kasugamycin and the allicin have unstable chemical properties under alkaline conditions. (2) The formula includes no pH adjusting agent but the pH of the formula is finally adjusted to 6-7. Thus, the pH adjustment method is not clear, indicating that the technical solution thereof is incomplete. (3) Sodium butyl naphthalene sulfonate as a surfactant only takes up 1-3% in the formulation, which can hardly ensure that oily allicin is desirably emulsified and becomes transparent when dissolved in water. Thus, there is a problem with the preparation in physical stability. (4) Both sodium butyl naphthalene sulfonate and CMC-Na are anionic surfactants with relatively large anionic groups. The kasugamycin is easy to react with acidic substances to form a salt, increasing its solubility. However, it forms an insoluble matter when combined with weakly acidic anions. (5) The patent application includes ethanol in its technical solution, which will significantly reduce solubility of kasugamycin.
In view of the above existing technical problems, the present disclosure provides a biological pesticide preparation containing allicin and kasugamycin, and a preparation method and use thereof.
The objective of the disclosure can be achieved by the following technical solutions.
A biological pesticide microemulsion for controlling rice diseases, including the following components:
1-50 parts of active ingredients, 15-30 parts of emulsifier, 1-10 parts of synergist, pH adjusting agent, and water as balance to a total of 100%; where the active ingredients are allicin and kasugamycin in a mass ratio of (1-80):(1-80) and preferably (1-20):(1-20).
In the above microemulsion, the emulsifier may be one or a combination of two or more of nonylphenol polyoxyethylene ether phosphate, tristyrylphenol polyoxyethylene ether, alkylphenol formaldehyde resin polyoxyethylene ether, dibenzyl biphenyl polyoxyethylene ether, castor oil polyoxyethylene ether, AEO-15, Tween 80, Tween 60, ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether and iso-tridecanol polyoxyethylene ether; the synergist may be one or a combination of two or more of lauryl hydroxypropyl phosphate betaine, betaine, erucamide propyl hydroxysultaine, polyether modified polyorganosiloxane, lecithin, and azone; and the pH adjusting agent may be one or a combination of two or more of citric acid, acetic acid, phosphoric acid and hydrochloric acid.
In some preferred technical solutions:
the emulsifier in the microemulsion may be at least one of Tween 80, alkylphenol formaldehyde resin polyoxyethylene ether, AEO-15 and tristyryl phenol polyoxyethylene ether; the synergist in the microemulsion may be at least one of lauryl hydroxypropyl phosphate betaine, lecithin, and azone.
A method for preparing the above biological pesticide microemulsion, including: mixing allicin, emulsifier and synergist uniformly, adding water and pH adjusting agent, stirring uniformly, finally adding kasugamycin and stirring uniformly to obtain the biological pesticide microemulsion.
A biological pesticide oil in water (O/W) emulsion for controlling rice diseases, including the following components:
1-50 parts of active ingredients, 3-15 parts of emulsifier, 1-10 parts of synergist, 0.01-5 parts of antifreezing agent, 0.01-0.5 parts of thickener, pH adjusting agent, and water as balance to a total of 100%; where the active ingredients are allicin and kasugamycin in a mass ratio of (1-80):(1-80) and preferably (1-20):(1-20).
In the above O/W emulsion, the emulsifier may be one or a combination of two or more of nonylphenol polyoxyethylene ether phosphate, tristyrylphenol polyoxyethylene ether, alkylphenol formaldehyde resin polyoxyethylene ether, dibenzyl biphenyl polyoxyethylene ether, castor oil polyoxyethylene ether, fatty alcohol polyoxyethylene ether, Tween 80, Tween 60, ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether and iso-tridecanol polyoxyethylene ether;
the synergist may be one or a combination of two or more of lauryl hydroxypropyl phosphate betaine, betaine, erucamide propyl hydroxysultaine, polyether modified polyorganosiloxane, lecithin, and azone;
the antifreezing agent may be one or a combination of two or more of ethylene glycol, glycerol and propylene glycol;
the thickener may be one or a combination of two of xanthan gum and polyvinyl alcohol 1788; and the pH adjusting agent may be one or a combination of two or more of citric acid, acetic acid, phosphoric acid and hydrochloric acid.
In some preferred technical solutions:
the emulsifier in the O/W emulsion may be at least one of ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether, Tween 60 and castor oil polyoxyethylene ether; and
the synergist in the O/W emulsion may be lauryl hydroxypropyl phosphate betaine or betaine.
A method for preparing the above biological pesticide O/W emulsion for controlling rice diseases, including: stirring allicin, emulsifier, and synergist uniformly to obtain an oil phase, adding pH adjusting agent, antifreezing agent and thickener to water and stirring uniformly to obtain a water phase, adding the oil phase to the water phase under high-speed shearing conditions, and shearing uniformly to obtain the O/W emulsion.
A technical solution of the present disclosure provides use of the biological pesticide microemulsion in preparation of medicines for controlling rice blast, rice bacterial leaf blight and rice panicle blight.
A technical solution of the present disclosure provides use of the biological pesticide O/W emulsion in preparation of medicines for controlling rice blast, rice bacterial leaf blight and rice panicle blight.
The present disclosure has the following beneficial effects:
In the present disclosure, the biological pesticide preparation innovatively takes the form of microemulsion or O/W emulsion, with an optimized formula including a pH adjusting agent to enhance stability of kasugamycin. Moreover, the present disclosure uses no organic solvent, and includes a synergist during preparation which effectively improves efficacy of the product. The biological pesticide preparation can be used to control rice diseases, and has outstanding effects on rice blast, rice bacterial leaf blight and rice panicle blight, which broadens the application range of the product with an excellent effect.
The present disclosure is further described below with reference to the embodiments, but the protection scope of the present disclosure is not limited herein.
Example 1
21% allicin-kasugamycin microemulsion 20+1
A biological pesticide microemulsion for rice disease included the following components by weight: 20 kg of allicin, 1 kg of kasugamycin, 8 kg of Tween 80, 22 kg of styryl phenyl polyoxyethylene ether, 5 kg of lauryl hydroxypropyl phosphate betaine, 0.3 kg of citric acid and water as balance to a total weight of 100 kg.
The microemulsion was prepared as follows. The allicin, the Tween 80, the styryl phenyl polyoxyethylene ether and the lauryl hydroxypropyl phosphate betaine were added to a stirring tank and stirred uniformly. The water was added and stirred uniformly. Then the citric acid was added and stirred uniformly. The kasugamycin was added and stirred to achieve complete dissolution to obtain the microemulsion.
Example 2
44% allicin-kasugamycin O/W emulsion 40+4
A biological pesticide O/W emulsion for rice disease included the following components by weight: 40 kg of allicin, 4 kg of kasugamycin, 2 kg of ethylene oxide propylene oxide block polyether (BASF Corporation, PE10500), 6 kg of cardanol polyoxyethylene ether (Shijiazhuang Haisen Chemical, GS7810), 3 kg of betaine, 1 kg of PVA1788, 5 kg of ethylene glycol, 0.3 kg of citric acid and water as balance to a total weight of 100 kg.
The O/W emulsion was prepared as follows. The allicin, 4 kg of PE10500, and betaine were added to a shearing tank and stirred uniformly. The water was added under high-speed shearing conditions and sheared for 1.5 h to achieve completely uniform shearing. The propylene glycol and the citric acid were added at one time and sheared uniformly. The kasugamycin was added and sheared uniformly. Then, the thickener PVA1788 was added and sheared uniformly to obtain the O/W emulsion. During preparation, high-speed shearing was carried out at 3,000 r/min by a shearing machine.
Example 3
12% allicin-kasugamycin microemulsion 10+2
A biological pesticide microemulsion for rice disease included the following components by weight: 10 kg of allicin, 2 kg of kasugamycin, 25 kg of Tween 80, 3 kg of azone, 0.3 kg of citric acid and water as balance to a total weight of 100 kg.
The microemulsion was prepared as follows. The allicin, the Tween 80 and the azone were added to a stirring tank and stirred uniformly. The water was added and stirred uniformly. Then the citric acid was added and stirred uniformly. The kasugamycin was added and stirred to achieve complete dissolution to obtain the microemulsion.
Example 4
24% allicin-kasugamycin O/W emulsion 20+4
A biological pesticide O/W emulsion for rice disease included the following components by weight: 20 kg of allicin, 4 kg of kasugamycin, 3 kg of ethylene oxide propylene oxide block polyether (BASF Corporation, PE10500), 6 kg of Tween 60, 5 kg of lauryl hydroxypropyl phosphate betaine, 2 kg of propylene glycol, 0.1 kg of xanthan gum, 0.3 kg of citric acid and water as balance to a total weight of 100 kg.
The O/W emulsion was prepared as follows. The allicin, the PE10500, and the lauryl hydroxypropyl phosphate betaine were added to a shearing tank and stirred uniformly. The water was added under high-speed shearing conditions and sheared for 1.5 h to achieve completely uniform shearing. The propylene glycol and the citric acid were added at one time and sheared uniformly. The kasugamycin was added and sheared uniformly. Then, the thickener xanthan gum was added and sheared uniformly to obtain the O/W emulsion. During preparation, high-speed shearing was carried out at 3,000 r/min by a shearing machine.
Example 5
20% allicin-kasugamycin O/W emulsion 10+10
A biological pesticide O/W emulsion for rice disease included the following components by weight: 10 kg of allicin, 10 kg of kasugamycin, 2 kg of ethylene oxide propylene oxide block polyether (BASF Corporation, PE10500), 3 kg of lauryl hydroxypropyl phosphate betaine, 5 kg of ethylene glycol, 0.1 kg of xanthan gum, 0.3% of citric acid and water as balance to a total weight of 100 kg.
The O/W emulsion was prepared as follows. The allicin, the PE10500, and the lauryl hydroxypropyl phosphate betaine were added to a shearing tank and stirred uniformly. The water was added under high-speed shearing conditions and sheared for 1.5 h to achieve completely uniform shearing. The ethylene glycol and the citric acid were added at one time and sheared uniformly. The kasugamycin was added and sheared uniformly. Then, the thickener xanthan gum was added and sheared uniformly to obtain the O/W emulsion. During preparation, high-speed shearing was carried out at 3,000 r/min by a shearing machine.
Example 6
10% allicin-kasugamycin microemulsion 5+5
A biological pesticide microemulsion for rice disease included the following components by weight: 5 kg of allicin, 5 kg of kasugamycin, 20 kg of alkylphenol formaldehyde resin polyoxyethylene ether, 4 kg of azone, 0.3% of citric acid and water as balance to a total weight of 100 kg.
The microemulsion was prepared as follows. The allicin, the alkylphenol formaldehyde resin polyoxyethylene ether and the azone were added to a stirring tank and stirred uniformly. The water was added and stirred uniformly. Then the citric acid was added and stirred uniformly. The kasugamycin was added and stirred to achieve complete dissolution to obtain the microemulsion.
Example 7
6% allicin-kasugamycin microemulsion 1+5
A biological pesticide microemulsion for rice disease included the following components by weight: 1 kg of allicin, 5 kg of kasugamycin, 15 kg of alkylphenol formaldehyde resin polyoxyethylene ether, 3 kg of lauryl hydroxypropyl phosphate betaine, 3 kg of azone, 0.3% of citric acid and water as balance to a total weight of 100 kg.
The microemulsion was prepared as follows. The allicin, the alkylphenol formaldehyde resin polyoxyethylene ether, the lauryl hydroxypropyl phosphate betaine and the azone were added to a stirring tank and stirred uniformly. The water was added and stirred uniformly. Then the citric acid was added and stirred uniformly. The kasugamycin was added and stirred to achieve complete dissolution to obtain the microemulsion.
Example 8
11% allicin-kasugamycin O/W emulsion 1+10
A biological pesticide O/W emulsion for rice disease included the following components by weight: 1 kg of allicin, 10 kg of kasugamycin, 5 kg of GS7807 (cardanol polyoxyethylene ether), 5 kg of betaine, 5 kg of glycerol, 2 kg of PVA1788, 0.3% of citric acid and water as balance to a total weight of 100 kg.
The O/W emulsion was prepared as follows. The allicin, the GS7807 (cardanol polyoxyethylene ether), and the betaine were added to a shearing tank and stirred uniformly. The water was added under high-speed shearing conditions and sheared for 1.5 h to achieve completely uniform shearing. The citric acid was added and sheared uniformly. The kasugamycin was added and sheared uniformly. Then, the thickener PVA1788 was added and sheared uniformly to obtain the O/W emulsion. During preparation, high-speed shearing was carried out at 3,000 r/min by a shearing machine.
Example 9
21% allicin-kasugamycin microemulsion 1+20
A biological pesticide microemulsion for rice disease included the following components by weight: 1 kg of allicin, 20 kg of kasugamycin, 15 kg of AEO-15, 5 kg of lauryl hydroxypropyl phosphate betaine, 1 kg of lecithin, 0.3 kg of citric acid and water as balance to a total weight of 100 kg.
The microemulsion was prepared as follows. The allicin, the AEO-15, the lauryl hydroxypropyl phosphate betaine and the lecithin were added to a stirring tank and stirred uniformly. The water was added and stirred uniformly. Then the citric acid was added and stirred uniformly. The kasugamycin was added and stirred to achieve complete dissolution to obtain the microemulsion.
Comparative Example 1
14% allicin-kasugamycin soluble liquid: 6 kg of allicin, 8 kg of kasugamycin, 1 kg of sodium butyl naphthalene sulfonate, and 3 kg of CMC-Na were added to 82 kg of water and stirred uniformly.
Comparative Example 2
14% allicin-kasugamycin soluble powder: 5 kg of allicin, 9 kg of kasugamycin, 1 kg of sodium butyl naphthalene sulfonate, and 3 kg of CMC-Na were added to 82 kg of water and stirred uniformly.
The present disclosure will be described in detail below with reference to application embodiments to facilitate further understanding of the present disclosure.
Example 10
The following experiment was carried out to determine effects of different products prepared with allicin and kasugamycin in controlling rice blast. Specific implementations were as follows.
Test pesticides: 21% allicin-kasugamycin microemulsion (20:1), 44% allicin-kasugamycin O/W emulsion, 12% allicin-kasugamycin microemulsion, 24% allicin-kasugamycin O/W emulsion, 20% allicin-kasugamycin O/W emulsion, 10% allicin-kasugamycin microemulsion, 6% allicin-kasugamycin microemulsion, 11% allicin-kasugamycin O/W emulsion, and 21% allicin-kasugamycin microemulsion (1:20).
Control agents: 2% kasugamycin aqueous solution, 5% allicin microemulsion, (Comparative Example 1) 14% allicin-kasugamycin soluble liquid, and (Comparative Example 2) 14% allicin-kasugamycin soluble powder.
Control object: rice blast
Environmental conditions: the experiment was carried out in Hongze County, Huai'an City in China, and conditions for cultivation, fertilizer and water management and the like were consistent among experimental plots.
Experimental design: the experiment had 14 treatments with 4 repeats a treatment, and a total of 56 plots with 20 square meters each plot. The plots were randomly arranged. The 14 treatments were:
(1) 21% allicin-kasugamycin microemulsion (20:1), (2) 44% allicin-kasugamycin O/W emulsion, (3) 12% allicin-kasugamycin microemulsion, (4) 24% allicin-kasugamycin O/W emulsion, (5) 20% allicin-kasugamycin O/W emulsion, (6) 10% allicin-kasugamycin microemulsion, (7) 6% allicin-kasugamycin microemulsion, (8) 11% allicin-kasugamycin O/W emulsion, (9) 21% allicin-kasugamycin microemulsion (1:20), (10) 14% allicin-kasugamycin soluble liquid, (11) 14% allicin-kasugamycin soluble powder, (12) 2% kasugamycin aqueous solution, (13) 5% allicin microemulsion, and (14) water as control.
Experimental method: in the experiment, agents were applied once at an initial stage of rice breakage, and once again at a full heading stage. 1.5 kg of water was added in application for each plot. Spraying was carried out with a 3WBD-16B knapsack electric sprayer with a single metal nozzle and adjusted spray speed, leaving no residual liquid.
Investigation method: 20 d after application, when panicle neck blast disease in the control area was stable with obvious symptoms, sampling was carried out at 5 points in each plot, with 50 rice plants at each point to investigate incidence of rice disease. A total number of panicles investigated in each plot, number of diseased panicles and disease level were recorded to calculate disease index and control effect.
Classification was carried out based on disease status according to the following standards. The disease index and relative control effect were calculated according to formula.
Level 0: no disease;
Level 1: less than 5% of loss per panicle (few branches were affected);
Level 3: 6%-20% of loss per panicle (about one-third of branches were affected);
Level 5: 21%-50% of loss per panicle (panicle neck or main cob was affected, and grain was half-deflated);
Level 7: 51%- 7 0% of loss per panicle (panicle neck was affected, and most grains were collapsed);
Level 9: 71%-100% of loss per panicle (cob was affected, causing white panicle)
Disease index=XE(number of diseased plants in each levelxrelative level number)X100 total number of plants investigatedx9
Controleffect=disease index of control-disease index of treatmentX100 disease index of control
Results were shown in Table 1.
Table 1 Control effect of test agents on rice blast
Dosage of active Disease Control effect Treatment ingredient (g/hm 2) index (%)
21% allicin-kasugamycin microemulsion (20:1) Example 1 45 1.94 83.93c
44% allicin-kasugamycin O/W emulsion 45 1.87 84.56c Example 2
12% allicin-kasugamycin microemulsion 45 1.53 87.33b Example 3
24% allicin-kasugamycin O/W emulsion 45 1.63 86.50b Example 4
20% allicin-kasugamycin O/W emulsion 45 1.27 89.53a Example 5
10% allicin-kasugamycin microemulsion 45 1.17 90.36a Example 6
6% allicin-kasugamycin microemulsion 45 1.09 91.00a Example 7
11% allicin-kasugamycin O/W emulsion 45 1.50 81.59b Example 8
21% allicin-kasugamycin microemulsion (1:20) Example 9 45 1.58 80.96b
(Comparative Example 1) 14% allicin-kasugamycin soluble liquid 45 2.38 80.35e
(Comparative Example 2) 14% allicin-kasugamycin soluble powder 45 2.16 82.19d
2% kasugamycin aqueous solution 45 2.59 78.60f
5% allicin microemulsion 45 4.52 62.63g
in
Blank control 0 12.10
Note: comparison was carried out with Duncan's new multiple range method with the DPS software, where a lowercase alphabet indicated 5% significance.
Experimental results showed that, control effects of combinations of allicin and kasugamycin on rice blast were significantly higher than that of a single agent of kasugamycin or allicin. Compared with the Comparative Examples, each Example had a higher control effect on rice blast, which can reduce dosage and cost of agents. Therefore, use of the allicin in combination with the kasugamycin in production practice for control of rice blast was economical, efficient, and environmentally friendly.
Example 11
Rice blast fungus was collected from rice fields in Hongze County, Huai'an City in China. It was isolated from a diseased rice panicle, identified, purified by a monospore method, and stored with potato dextrose agar (PDA) medium for later use.
Test agent: 70% kasugamycin technical material, 50% allicin technical concentrate
Determination of virulence of test agent on rice blast fungus
The rice blast fungus was inoculated on an oatmeal-tomato juice-agar medium and cultured at °C for 7 d. The hyphae and conidia were gently washed off with sterile water, and a suspension thereof was uniformly spread on medium in a new plate with about 400 L each dish, and blown dry on an ultra-clean workbench. Cultivation was carried out in an incubator at 25°C for about 40 h until a layer of sparse aerial mycelium grew on a surface of the medium. A small amount of sterile water was added to colonies. The aerial hyphae on the surface were gently wiped off with a cotton swab. The hyphae and conidia on the surface of the medium were washed with water. After drying naturally, 2 layers of gauze were used to cover the surface of the medium and cultivation was carried out at °C under light conditions until a large number of conidia were produced. The conidia were washed off from the medium and centrifuged. A supernatant was discarded. Precipitated conidia were suspended in a 0.05% Tween 80 aqueous solution, counted on a hemocytometer. A concentration of spore suspension was adjusted to 2x10 5/ml.
The kasugamycin technical material and the allicin technical concentrate were dissolved with methanol into 1,000 mg/L stock solutions, and combined in a ratio of allicin to kasugamycin of 50:1, :1, 30:1, 20:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:20, 1:30, 1:40, and 1:50. The prepared agents were diluted with sterile water in a constant ratio to obtain a series of concentrations. 5 concentrations were used in each experiment, with clean water as control.
4 pots were treated for each concentration, with 10 plants per pot. The pots were filled with water, and then the rice seedlings were sprayed with a spray tower quantitatively. 4 pots for the same concentration were sprayed together at the center of rotary table of the spray tower with 20 ml of agent. The top and side nozzles were used when applying agents. After spraying, the water in the pots was poured to prevent part of the agent sprayed into the pot from being absorbed by the rice and affecting measurement results. At 24 h after spraying, a glass sprayer for thin layer chromatography was used to inoculate spores of rice blast fungus. Then the pots were moved into a moisturizing cover, and kept in the dark at 26°C for 24 h with RH 100% and a water film on leaves. The pots were then moved to a plant growth room and cultivated with a cycle of 12 h lighting at 10,000 LUX and 12h in the dark and RH 85%-90% at 26-28°C for 7 d. Results were investigated. Disease levels of the second and the third complete leaves were recorded.
Classification of disease was carried out based on the following standards (with leaf as unit). The disease index and relative control effect were calculated according to formula.
Level 0: the whole plant not affected;
Level 1: brown spots;
Level 3: typical spindle lesions accounting for less than 5% of leaf area;
Level 5: typical lesions accounting for 6-25% of leaf area;
Level 7: typical lesions accounting for 26-50% of leaf area;
Level 9: typical lesions accounting for more than 50% of leaf area.
Disease index= E(number of diseased plants in each levelxrelative level number) X 100 total number of plants investigatedx9
Control effect=disease index of control-disease index of treatment x 100 disease index of control
Data were processed with the DPS software to obtain a regression equation and EC50. Synergistic effects of compounding agents were calculated according to Wadley's method. Namely, SR<0.5 indicated antagonism when the two agents were combined, 0.5<SR<1.5 indicated an additive effect when the two agents were combined, and SR>1.5 indicated a synergistic effect when the two agents were combined.
Experimental results were shown in Table 2.
Table 2 Determination results of virulence of test agent on rice blast fungus
EC5o Agent Regression equation ofvirulence Correlation coefficient (r) Synergy coefficient (SR) (pg/mL)
Allicin (A) y=3.0828+2.0059x 9.0328 0.9955
Kasugamycin (B) y=4.9507+1.4387x 1.0821 0.9910
A:B= 50:1 y=3.6091+1.4752x 8.7672 0.9828 0.9006
A:B= 40:1 y=3.6644+1.5866x 6.9471 0.9949 1.1026
A:B= 30:1 y=3.8665+1.5688x 5.2787 0.9991 1.3833
A:B=20:1 y=3.4627+2.8490x 3.4640 0.9959 1.9317
A:B=10:1 y=3.8679+2.6532x 2.6712 0.9953 2.0274
A:B=5:1 y=4.5904+2.3102x 1.5042 0.9891 2.6994
A:B=1:1 y=5.0390+1.4543x 0.9402 0.9977 2.0556
A:B=1:5 y=5.5518+2.3715x 0.5852 0.9985 2.1670
A:B=1:10 y=5.6112+2.4568x 0.5639 0.9956 2.0859
A:B=1:20 y-5.5332+2.4632x 0.6075 0.9923 1.8592
A:B= 1:30 y=5.0821+1.4554x 0.8783 0.9964 1.2680
A:B= 1:40 y=5.0473+1.4332x 0.9268 0.9982 1.1932
A:B= 1:50 y=4.9959+1.4870x 1.0064 0.9968 1.0941
A ratio range was A:B=50:1 to 1:50, preferably A:B=20:1 to 1:20.
It can be seen from Table 2 that, combination of the allicin and the kasugamycin in a ratio of 20:1 to 1:20 had a significantly increased effect on rice blast fungus. This indicated that, combination of the two agents had a significant synergistic effect on rice blast.
Example 12
The following experiment was carried out to determine effects of different products prepared with allicin and kasugamycin in controlling rice bacterial leaf blight. Specific implementations were as follows.
Test pesticides: 21% allicin-kasugamycin microemulsion (20:1), 44% allicin-kasugamycin O/W emulsion, 12% allicin-kasugamycin microemulsion, 24% allicin-kasugamycin O/W emulsion, 20% allicin-kasugamycin O/W emulsion, 10% allicin-kasugamycin microemulsion, 6% allicin-kasugamycin microemulsion, 11% allicin-kasugamycin O/W emulsion, and 21% allicin-kasugamycin microemulsion (1:20).
Control agents: 2% kasugamycin aqueous solution, 5% allicin microemulsion, (Comparative Example 1) 14% allicin-kasugamycin soluble liquid, and (Comparative Example 2) 14% allicin-kasugamycin soluble powder.
Control object: rice bacterial leaf blight
Environmental conditions: the experiment was carried out in Hongze County, Huai'an City in China, and conditions for cultivation, fertilizer and water management and the like were consistent among experimental plots.
Experimental design: the experiment had 14 treatments with 4 repeats a treatment, and a total of 56 plots with 20 square meters each plot. The plots were randomly arranged. The 14 treatments were:
(1) 21% allicin-kasugamycin microemulsion (20:1), (2) 44% allicin-kasugamycin O/W emulsion, (3) 12% allicin-kasugamycin microemulsion, (4) 24% allicin-kasugamycin O/W emulsion, (5) 20% allicin-kasugamycin O/W emulsion, (6) 10% allicin-kasugamycin microemulsion, (7) 6% allicin-kasugamycin microemulsion, (8) 11% allicin-kasugamycin O/W emulsion, (9) 21% allicin-kasugamycin microemulsion (1:20), (10) 14% allicin-kasugamycin soluble liquid, (11) 14% allicin-kasugamycin soluble powder, (12) 2% kasugamycin aqueous solution, (13) 5% allicin microemulsion, and (14) water as control.
Experimental method: in the experiment, agents were applied once at an initial stage of rice bacterial leaf blight, and once again at an interval of 7 d, for a total of 2 times. 1.5 kg of water was added in application for each plot. Spraying was carried out with a 3WBD-16B knapsack electric sprayer with a single metal nozzle and adjusted spray speed, leaving no residual liquid.
Investigation method: at day 14 after the final application, when rice bacterial leaf blight in the control area was stable with obvious symptoms, sampling was carried out at 5 points in each plot diagonally with 50 rice plants at each point to investigate incidence of rice disease on flag leaf and other two leaves below the flag leaf. A total number of leaves investigated in each plot, number of diseased leaves and disease level were recorded to calculate disease index and control effect.
Classification was carried out based on disease status according to the following standards. The disease index and relative control effect were calculated according to formula.
Level 0: no disease;
Level 1: lesions accounting for less than 10% of leaf area;
Level 3: lesions accounting for 11-25% of leaf area;
Level 5: lesions accounting for 26-45% of leaf area;
Level 7: lesions accounting for 46-65% of leaf area;
Level 9: lesions accounting for more than 65% of leaf area.
Disease index=XE(number of diseased leaf in each levelxrelative level number)X100 total number of leaves investigatedx9
Control effect=disease index of control-disease index of treatment X 100 disease index of control
Results were shown in Table 3.
Table 3 Control effects of test agents on rice bacterial leaf blight Dosage of active Disease Control effect Treatment ingredient (g/hm 2) index (%)
21% allicin-kasugamycin microemulsion (20:1) 45 1.11 82.75d Example 1
44% allicin-kasugamycin O/W emulsion 45 0.99 84.65c Example 2
12% allicin-kasugamycin microemulsion 45 0.82 87.30b Example 3
24% allicin-kasugamycin O/W emulsion 45 0.83 87.06b Example 4
20% allicin-kasugamycin O/W emulsion 45 0.68 89.43a Example 5
10% allicin-kasugamycin microemulsion 45 0.64 90.13a Example 6
6% allicin-kasugamycin microemulsion 45 0.86 86.60b Example 7
11% allicin-kasugamycin O/W emulsion 45 0.97 84.93c Example 8
21% allicin-kasugamycin microemulsion (1:20) Example 9 45 1.14 82.25d
(Comparative Example 1) 14% allicin-kasugamycin soluble liquid 45 1.24 80.69e
(Comparative Example 2) 14% allicin-kasugamycin soluble powder 45 1.28 80.19e
2% kasugamycin aqueous solution 45 2.04 68.30g
5% allicin microemulsion 45 1.53 76.26f
Blank control 0 6.44
Note: comparison was carried out with Duncan's new multiple range method with the DPS software, where a lowercase alphabet indicated 5% significance.
Experimental results showed that, control effects of combinations of allicin and kasugamycin on rice bacterial leaf blight were significantly higher than that of a single agent of kasugamycin or allicin. Compared with the Comparative Examples, each Example had a higher control effect on rice bacterial leaf blight, which can reduce dosage and cost of agents. Therefore, use of the allicin in combination with the kasugamycin in production practice for control of rice bacterial leaf blight was economical, efficient, and environmentally friendly.
Example 13
Xanthomonas oryzae was collected from rice fields in Hongze County, Huai'an City in China. It was isolated from a diseased paddy rice leaf, identified, vacuum freeze dried, and stored in a refrigerator at -18°C. Before measurement, streaking was carried out on nutrient agar (NA) solid medium to pick a single colony which was then streaked and cultured. Then, inoculation was carried out with nutrient broth (NB) medium and culturing was carried out under shaking for 24 h. A bacterial suspension with a number of 20 as shown by a turbidimeter was prepared with sterile water for later use.
Test agent: 70% kasugamycin technical material, 50% allicin technical concentrate
Determination of virulence of test agent on Xanthomonas oryzae
The kasugamycin technical material and the allicin technical concentrate were dissolved with methanol into 1,000 mg/L stock solutions, and combined in a ratio of allicin to kasugamycin of 50:1, :1, 30:1, 20:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:20, 1:30, 1:40, and 1:50. The prepared agents were diluted with sterile water in a constant ratio to obtain a series of concentrations. 5 concentrations were used in each experiment. 25 ml of agent-containing NB medium was added to a 50 ml empty Erlenmeyer flask. Then 100 L of bacterial suspension was inoculated. Treatment with clean water was used as control. 4 flasks were used for repetition for each concentration.
A turbidity method was used. After culturing under shaking at 28°C and 175 rpm for 36 h, turbidity was measured to calculate inhibition rate of each agent on pathogen growth. Data were processed with the DPS software to obtain a regression equation and EC50. Synergistic effects of compounding agents were calculated according to Wadley's method. Namely, SR<0.5 indicated antagonism when the two agents were combined, 0.5<SR<1.5 indicated an additive effect when the two agents were combined, and SR>1.5 indicated a synergistic effect when the two agents were combined.
Experimental results were shown in Table 4.
Table 4 Determination results of virulence of test agent on Xanthomonas oryzae EC5o Agent Regression equation ofvirulence Correlation coefficient (r) Synergy coefficient (SR) (pg/mL)
Allicin (A) y=4.3322+1.4790x 2.8283 0.9976
Kasugamycin (B) y=4.0587+1.4712x 4.3635 0.9938
A:B= 50:1 y=4.3311+1.5093x 2.7746 0.9969 1.0264
A:B= 40:1 y=4.3265+1.5637x 2.6961 0.9977 1.0581
A:B= 30:1 y=4.4808+1.4053x 2.3413 0.9955 1.2219
A:B=20:1 y=4.8163+1.4459x 1.3398 0.9963 2.1470
A:B=10:1 y=4.7891+1.5080x 1.3799 0.9922 2.1174
A:B=5:1 y=4.8671+1.5285x 1.2217 0.9931 2.4593
A:B-1:1 y=4.7864+1.5662x 1.3690 0.9960 2.5070
A:B-1:5 y=4.7021+1.3834x 1.6420 0.9965 2.4370
A:B-1:10 y=4.4984+1.4893x 2.1716 0.9944 1.9149
A:B-1:20 y=4.5756+1.3911x 2.0189 0.9967 2.1069
A:B= 1:30 y=4.0835+1.6311x 3.6467 0.9902 1.1760
A:B= 1:40 y=4.0985+1.6498x 3.5193 0.9907 1.2237
A:B= 1:50 y=3.9785+1.5196x 4.7012 0.9940 0.9184
A ratio range was A:B=50:1 to 1:50, preferably A:B=20:1 to 1:20.
It can be seen from Table 4 that, combinations of the allicin and the kasugamycin in a ratio of :1 to 1:20 had a significantly increased effect on Xanthomonas oryzae. This indicated that, combination of the two agents had a significant synergistic effect on rice bacterial leaf blight.
Example 14
The following experiment was carried out to determine effects of different products prepared with allicin and kasugamycin in controlling rice panicle blight. Specific implementations were as follows.
Burkholderia glumae was collected from rice fields in Fuyang District, Hangzhou city in China. It was isolated from a rice panicle with symptoms of rice panicle blight and identified. Before measurement, streaking was carried out on NA solid medium to pick a single colony which was then streaked and cultured. Then, inoculation was carried out with NB medium and culturing was carried out under shaking for 24 h. A 10 8/ml bacterial suspension was prepared with sterile water for later use.
Test pesticides: 21% allicin-kasugamycin microemulsion (20:1), 44% allicin-kasugamycin O/W emulsion, 12% allicin-kasugamycin microemulsion, 24% allicin-kasugamycin O/W emulsion, 20% allicin-kasugamycin O/W emulsion, 10% allicin-kasugamycin microemulsion, 6% allicin-kasugamycin microemulsion, 11% allicin-kasugamycin O/W emulsion, and 21% allicin-kasugamycin microemulsion (1:20).
Control agents: 2% kasugamycin aqueous solution, 5% allicin microemulsion, (Comparative Example 1) 14% allicin-kasugamycin soluble liquid, and (Comparative Example 2) 14% allicin-kasugamycin soluble powder.
Experimental design: a pot cultivating method was used. A total of 14 treatments with 9 test agents, 4 control agents and water control were included in the experiment, with 4 pots each treatment and 10 plants each pot. Based on a total active ingredient dosage of 45 g/hm2 and 50 kg of water per mu, a concentration of test agent was set to 60 mg a.i./L. The pots were filled with water 5 d before rice hearing, and then the rice seedlings were sprayed with a spray tower quantitatively. 4 pots for the same agent were sprayed together at the center of rotary table of the spray tower with 20 ml of agent. The top and side nozzles were used when applying agents. After spraying, the water in the pots was poured to prevent part of the agent sprayed into the pot from being absorbed by the rice and affecting measurement results. After drying naturally for 24 h, the prepared 108cfu/ml Burkholderia glumae suspension was sprayed and inoculated in an average amount of 10 ml on each rice plant. Then the pots were moved into a moisturizing cover, and kept in the dark at 26°C for 24 h with RH 100% and a water film on leaves. The pots were then moved to a plant growth room and cultivated with a cycle of 12 h lighting at 10,000 LUX and 12h in the dark and RH 85%-90% at 26-28°C. Agents were applied once again at an interval of 7 d. Cultivation was continued to a mature period. When disease was obvious with the control treatment, results were investigated.
Investigation method: when the disease of panicle blight was stable and the symptoms were obvious with water control treatment, disease of each rice plant and number of diseased grains per panicle were investigated. Total number of grains and number of diseased grains in each pot were recorded, and incidence and control effect were calculated.
Incidence= number of diseased grains X100 total number of grains investigated
Control effect= incidence of control-incidence of treatmentX100 incidence of control
Results were shown in Table 5.
Table 5 Control effects of test agents on rice panicle blight
Averagol Dosage of active AverageControl Treatment incidence rate ingredient (mg/kg) effect (%)
21% allicin -kasugamycin microemulsion (20:1) Example 1 60 8.40 84.72b
44% allicin-kasugamycin O/W emulsion 60 8.21 85.08b Example 2
12% allicin-kasugamycin microemulsion 60 7.52 86.34b Example 3
24% allicin-kasugamycin O/W emulsion 60 7.94 85.57b Example 4
20% allicin-kasugamycin O/W emulsion 60 6.19 88.75a Example 5
10% allicin-kasugamycin microemulsion 60 6.42 88.33a Example 6
6% allicin-kasugamycin microemulsion 60 7.86 85.71b Example 7
11% allicin-kasugamycin O/W emulsion 60 7.73 85.95b Example 8
21% allicin-kasugamycin microemulsion (1:20) Example 9 60 8.43 84.68b
(Comparative Example 1) 14% allicin-kasugamycin soluble liquid 60 9.60 82.56c
(Comparative Example 2) 14% allicin-kasugamycin soluble powder 60 9.99 81.83c
2% kasugamycin aqueous solution 60 17.40 68.37e
5% allicin microemulsion 60 13.66 75.17d
Blank control 0 55.02
Note: comparison was carried out with Duncan's new multiple range method with the DPS software, where a lowercase alphabet indicated 5% significance.
Experimental results showed that, control effects of combinations of allicin and kasugamycin on rice panicle blight were significantly higher than that of a single agent of kasugamycin or allicin. Compared with the Comparative Examples, each Example had a higher control effect on rice panicle blight, which can reduce dosage and cost of agents. Therefore, use of the allicin in combination with the kasugamycin in production practice for control of rice panicle blight was economical, efficient, and environmentally friendly.
Example 15
Burkholderia glumae was collected from rice fields in Fuyang District, Hangzhou city in China. It was isolated from a rice panicle with symptoms of rice panicle blight and identified. Before measurement, streaking was carried out on NA solid medium to pick a single colony which was then streaked and cultured. Then, inoculation was carried out with NB medium and culturing was carried out under shaking for 24 h. A 107 cfu/ml bacterial suspension was prepared with sterile water for later use.
Test agent: 70% kasugamycin technical material, 50% allicin technical concentrate
Determination of virulence of test agent on Burkholderiaglumae
The kasugamycin technical material and the allicin technical concentrate were dissolved with methanol into 1,000 mg/L stock solutions, and combined in a ratio of allicin to kasugamycin of 50:1, :1, 30:1, 20:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:20, 1:30, 1:40, and 1:50. The prepared agents were diluted with sterile water in a constant ratio to obtain a series of concentrations. 5 concentrations were used in each experiment. 25 ml of agent-containing NB medium was added to a 50 ml empty Erlenmeyer flask. Then 100 L of bacterial suspension was inoculated. Treatment with clean water was used as control. 4 flasks were used for repetition for each concentration.
A turbidity method was used. After culturing under shaking at 28°C and 175 rpm for 36 h, turbidity was measured to calculate inhibition rate of each agent on pathogen growth. Data were processed with the DPS software to obtain a regression equation and EC50. Synergistic effects of compounding agents were calculated according to Wadley's method. Namely, SR<0.5 indicated antagonism when the two agents were combined, 0.5<SR<1.5 indicated an additive effect when the two agents were combined, and SR>1.5 indicated a synergistic effect when the two agents were combined.
Experimental results were shown in Table 6.
Table 6 Determination results of virulence of test agent on Burkholderia glumae Agent Regression equation of EC5o Correlation Synergy coefficient virulence coefficient (r) (SR) (pg/mL)
Allicin (A) y=4.4158+1.6176x 2.2970 0.9880
Kasugamycin (B) y=3.9343+1.9158x 3.5997 0.9987
A:B= 50:1 y=4.4049+1.5020x 2.4901 0.9941 0.9290
A:B= 40:1 y=4.5105+1.4388x 2.1889 0.9985 1.0587
A:B= 30:1 y=4.6126+1.3970x 1.8936 0.9995 1.2274
A:B=20:1 y=4.8080+1.5587x 1.3280 0.9970 1.7600
A:B=10:1 y=4.9183+1.5285x 1.1310 0.9962 2.1000
A:B=5:1 y=4.9727+1.5059x 1.0426 0.9952 2.3446
A:B-1:1 y=4.7837+1.5544x 1.3776 0.9980 2.0358
A:B-1:5 y=4.7189+1.6756x 1.4715 0.9964 2.2350
A:B-1:10 y=4.6880+1.4061x 1.6668 0.9965 2.0538
A:B-1:20 y=4.5897+1.4824x 1.8913 0.9996 1.8532
A:B= 1:30 y=4.3297+1.3781x 3.0649 0.9937 1.1534
A:B= 1:40 y=4.1825+1.7395x 2.9511 0.9984 1.2031
A:B= 1:50 y=4.2392+1.4083x 3.4693 0.9943 1.0262
A ratio range was A:B=50:1 to 1:50, preferably A:B=20:1 to 1:20.
It can be seen from Table 6 that, combinations of the allicin and the kasugamycin in a ratio of :1 to 1:20 had a significantly increased effect on Burkholderia glumae. This indicated that, combination of the two agents had a significant synergistic effect on rice panicle blight.
The above embodiments are intended to merely explain and illustrate the technical solutions of the present disclosure, rather than to limit the scope of the present disclosure. It should be noted that several improvements and modifications may also be made by those of ordinary skill in the art without departing from the spirit of the present disclosure and fall within the scope of the claims of the present disclosure.
Claims (5)
1. A biological pesticide microemulsion for controlling rice diseases, comprising the following components:
1-50 parts of active ingredients, 15-30 parts of emulsifier, 1-10 parts of synergist, pH adjusting agent, and water as balance to a total of 100%; wherein the active ingredients are allicin and kasugamycin in a mass ratio of (1-80):(1-80) and preferably (1-20):(1-20).
2. The biological pesticide microemulsion for controlling rice diseases according to claim 1, wherein, the emulsifier is one or a combination of two or more of nonylphenol polyoxyethylene ether phosphate, tristyrylphenol polyoxyethylene ether, alkylphenol formaldehyde resin polyoxyethylene ether, dibenzyl biphenyl polyoxyethylene ether, castor oil polyoxyethylene ether, AEO-15, Tween , Tween 60, ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether and iso-tridecanol polyoxyethylene ether;
the synergist is one or a combination of two or more of lauryl hydroxypropyl phosphate betaine, betaine, erucamide propyl hydroxysultaine, polyether modified polyorganosiloxane, lecithin, and azone;and
the pH adjusting agent is one or a combination of two or more of citric acid, acetic acid, phosphoric acid and hydrochloric acid;
wherein,
the emulsifier is at least one of Tween 80, alkylphenol formaldehyde resin polyoxyethylene ether, AEO-15 and tristyryl phenol polyoxyethylene ether; and the synergist is at least one of lauryl hydroxypropyl phosphate betaine, lecithin, and azone.
3. A method for preparing the biological pesticide microemulsion according to claim 1, comprising: mixing allicin, emulsifier and synergist uniformly, adding water and pH adjusting agent, stirring uniformly, finally adding kasugamycin and stirring uniformly to obtain the biological pesticide microemulsion.
4. A biological pesticide oil in water (O/W) emulsion for controlling rice diseases, comprising the following components:
1-50 parts of active ingredients, 3-15 parts of emulsifier, 1-10 parts of synergist, 0.01-5 parts of antifreezing agent, 0.01-0.5 parts of thickener, pH adjusting agent, and water as balance to a total of 100%; wherein the active ingredients are allicin and kasugamycin in a mass ratio of (1-80):(1-80) and preferably (1-20):(1-20).
5. The biological pesticide O/W emulsion for controlling rice diseases according to claim 4, wherein,
the emulsifier is one or a combination of two or more of nonylphenol polyoxyethylene ether phosphate, tristyrylphenol polyoxyethylene ether, alkylphenol formaldehyde resin polyoxyethylene ether, dibenzyl biphenyl polyoxyethylene ether, castor oil polyoxyethylene ether, fatty alcohol polyoxyethylene ether, Tween 80, Tween 60, ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether and iso-tridecanol polyoxyethylene ether;
the synergist is one or a combination of two or more of lauryl hydroxypropyl phosphate betaine, betaine, erucamide propyl hydroxysultaine, polyether modified polyorganosiloxane, lecithin, and azone;
the antifreezing agent is one or a combination of two or more of ethylene glycol, glycerol and propylene glycol;
the thickener is one or a combination of two of xanthan gum and polyvinyl alcohol 1788; and
the pH adjusting agent is one or a combination of two or more of citric acid, acetic acid, phosphoric acid and hydrochloric acid;
wherein,
the emulsifier is at least one of ethylene oxide propylene oxide block polyether, cardanol polyoxyethylene ether, Tween 60 and castor oil polyoxyethylene ether; and
the synergist is lauryl hydroxypropyl phosphate betaine or betaine;
the method for preparing the biological pesticide O/W emulsion for controlling rice diseases comprise: mixing and stirring allicin, emulsifier, and synergist uniformly to obtain an oil phase, adding pH adjusting agent, antifreezing agent and thickener to water and stirring uniformly to obtain a water phase, adding the oil phase to the water phase under high-speed shearing conditions, and shearing uniformly to obtain the O/W emulsion.
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