Green pesticide based on glucose ester and derivatives thereof, preparation method and application

Abstract

The invention discloses a green pesticide based on glucose ester and derivatives thereof, and a preparation method and application thereof, and the green pesticide comprises the following steps: mixing one or more of glucose ester and derivatives thereof according to any proportion to obtain a first component, and compounding the first component with a second component to obtain a water-based pesticide formulation, such as an emulsion in water, a water suspending agent and the like; wherein the second component is a pesticide raw material selected from the group consisting of tebuconazole, zinc thiazole, thiediazole copper, colistin sulfate and validamycin. The green pesticide provided by the invention can achieve 90-100% of the prevention and treatment effect only by 1-40% of the application amount of commercially available pesticides, and solves the problems of high toxicity, difficult degradation and low utilization rate of the existing pesticides. The glucose ester and the derivative thereof are used as green pesticides, have the functions of dispersion and permeation promotion, have antibacterial activity, are green and efficient, are easily degraded by the environment, and can remarkably improve the pesticide efficiency and reduce the environmental influence.

Description

Technical Field

[0001] The present invention relates to the technical field of glycolipid biosurfactants, and in particular to a green pesticide based on glucose lipids and derivatives thereof, a preparation method and application thereof. Background Art

[0002] Agriculture is the key to the sustainable development of a country's economy, society and environment, and plant diseases are one of the important factors affecting agricultural production. According to the Food and Agriculture Organization of the United Nations (FAO), by 2050, the global population is expected to reach nearly 10 billion, which poses a huge challenge to food production. In order to meet the needs of the growing population, global agriculture needs to increase food production by 70% to 100% on the existing basis. However, the impact of plant diseases on crop yield and quality cannot be ignored. The losses caused by various plant diseases each year amount to hundreds of billions of dollars, which seriously affects the national economy and threatens the country's food security.

[0003] Plant diseases are mainly caused by pathogens such as fungi, bacteria, and viruses. The degree of crop damage ranges from mild symptoms to large-scale destruction of food crops, resulting in a significant reduction in agricultural production. Studies have shown that global crop diseases cause a loss of 10.1% to 28.1% in wheat production, 24.6% to 40.9% in rice, 19.5% to 41.1% in corn, and 11.0% to 32.4% in soybeans. 88% of the world's wheat is susceptible to stripe rust, resulting in an annual loss of 5.47 million tons of wheat production. Rice blast caused by rice blast alone has led to a 30% reduction in rice production worldwide, and these losses are enough to feed 60 million people. Bacterial diseases represented by rice streak disease and rice bacterial leaf blight are becoming increasingly serious, causing huge yield losses of 40% to 60% in agricultural production.

[0004] At present, pesticides are usually used to prevent and control plant diseases. The use of pesticides can effectively control crop diseases, insects, and weeds, and can save 30% to 40% of the total crop production losses in the world. However, traditional pesticides are mainly chemical pesticides, and their large-scale use not only puts tremendous pressure on the ecological environment, but also has an adverse impact on the quality and safety of agricultural products due to excessive pesticide residues. After entering the environment, pesticides also cause damage to the air, water, soil and biodiversity. With the accelerated elimination and banning of traditional highly toxic, high-risk, and inefficient pesticides, new green pesticides with high efficiency, low toxicity, and low residue have become the focus and mainstream trend of industry research and development, and pesticide formulations are developing in the direction of water-based, dust-free, controlled release and other efficient, safe, and green directions.

[0005] Most pesticide raw materials need to be processed to make commercial forms suitable for different occasions. In order to reduce the use of organic solvents, adjuvants are essential ingredients in pesticides. The development of safe, efficient, green and environmentally friendly pesticide adjuvants is one of the important directions for the greening of pesticides. Among them, surfactants have hydrophilic groups that are soluble in water and lipophilic groups that are soluble in oil. At low concentrations, they can significantly change the surface tension of liquids and keep the formulation stable for a long time. Pesticide products usually contain 1% to 10% of one or more surfactants, which help to disperse, emulsify, better diffuse or increase the contact area between the agent and the plant surface, so that the pesticide can penetrate better and improve the efficacy of the pesticide. However, most surfactants are currently chemically synthesized, which are not easily degraded in the environment and have certain ecotoxicity. In the context of circular economy and sustainable development, biosurfactants have attracted attention due to their good biocompatibility. They are usually obtained by microbial metabolic transformation and show low or non-toxic, biodegradable, and environmentally friendly characteristics. Therefore, it can completely replace chemical surfactants in pesticides and realize green pesticides.

[0006] Glucolipids (GLs) are a class of glycolipid biosurfactants and are also intermediates in the biosynthesis pathway of sophorolipids (SLs). Sophorolipids are produced by non-pathogenic yeast metabolism. The sugar head is composed of sophorose (two molecules of glucose bonded by β-1,2 glycosidic bonds). They are mainly divided into two categories: acid type and lactone type. They have certain insecticidal and antibacterial activities and can destroy cells by interfering with the rearrangement of cell membranes or lysing cell membranes. However, compared with traditional chemical antibacterial agents, their antibacterial activity is weaker, especially for fungi and Gram-negative bacteria, with a minimum inhibitory concentration of up to 1.5-15 g / L. In order to further enhance its antibacterial activity, a chemical modification strategy can be adopted to remove a glucose from the acid-type sophorolipid head group by deglycosylation to obtain glucose lipids. Glucose lipids are more hydrophobic, more surface active, and have better antibacterial activity than acid-type sophorolipids. Furthermore, through amide modification, various groups were grafted onto its carboxyl terminus to obtain glucose lipid derivatives, which exhibited excellent antibacterial activity and biocompatibility, and contributed to the green prevention and control of plant diseases.

[0007] In summary, with the development of agricultural modernization, the use of pesticides is becoming more and more common, but traditional pesticides often have environmental pollution, residue problems and impacts on non-target organisms. Therefore, the development of new environmentally friendly, safe and efficient green pesticides has become an urgent problem to be solved. Pesticide adjuvants are essential ingredients in pesticide formulations, with a content of up to 99%. The development of green and environmentally friendly pesticide adjuvants is a key factor in the green development of pesticides. Glucose lipids and their derivatives, as a biosurfactant, are considered to be an ideal candidate material due to their good biocompatibility and degradability. At the same time, the modified glucose lipids and their derivatives have better antibacterial activity and good surface / interfacial activity, which helps to reduce the application amount of pesticide originals and achieve the effect of "reducing application and increasing efficiency", providing new ideas for the creation of green pesticides for green prevention and control of plant diseases. Summary of the invention

[0008] The purpose of the present invention is to address the deficiencies of the prior art and provide a green pesticide based on glucose lipids and their derivatives, a preparation method and an application.

[0009] In a first aspect, an embodiment of the present invention provides a green pesticide based on glucose lipids and their derivatives, comprising:

[0010] One or more of glucose lipids and their derivatives are mixed in any proportion as a first component, and compounded with a second component to obtain a water-based pesticide formulation;

[0011] Wherein, the second component is selected from the pesticide technical including tebuconazole, zinc thiazole, thiophanate-copper, colistin sulfate, and validamycin.

[0012] Furthermore, when only glucose lipid is selected as the first component, the mass volume ratio of the glucose lipid in the water-based pesticide formulation is 1 to 40%.

[0013] Furthermore, when only glucose lipid derivatives are selected as the first component, the glucose lipid derivatives are glucose lipid D-arginine salt methyl ester derivatives, and the mass volume ratio of the glucose lipid derivatives in the water-based pesticide formulation is 1 to 80%.

[0014] Furthermore, when the water-based pesticide formulation is an emulsion in water, it includes, by mass volume ratio:

[0015] The mass volume ratio of pesticide technical is 5% to 25%, solvent is 15% to 40%, emulsifier is 5% to 15%, glucose lipid and its derivatives are 5% to 15%, other surfactants are 0% to 5%, and the rest is made up to 100% by water;

[0016] Wherein, the concentration of glucose lipid and its derivatives is 1-20g / L.

[0017] Further, the solvent is one or more of xylene, dichloromethane, cyclohexanone, dimethylformamide, alkylamides, and dimethyl sulfoxide mixed in any proportion;

[0018] The emulsifier is one or more of phosphate salts, polyoxyethylene ethers, and fatty alcohol polyethers mixed in any proportion;

[0019] Furthermore, when the water-based pesticide formulation is a water suspension, it includes, by mass volume ratio:

[0020] Pesticide technical 10% to 45%, glucose ester and its derivatives 1% to 20%, wetting and dispersing aid 4% to 10%, defoamer 0.1% to 0.5%, thickener 0.2% to 0.5%, antifreeze 1% to 3%, and the rest is made up to 100% by water;

[0021] Wherein, the concentration of glucose lipid and its derivatives is 1-20g / L.

[0022] Furthermore, the wetting and dispersing aid is one or more of alkylnaphthalene sulfates and lignin sulfonates mixed in any proportion;

[0023] The defoaming agent includes organic silicon defoaming agent, silicone oil defoaming agent, silicone defoaming agent;

[0024] The thickener is one or more of xanthan gum and magnesium aluminum silicate mixed in any proportion;

[0025] The antifreeze agent includes ethylene glycol, glycerol, propylene glycol, and polyvinyl alcohol.

[0026] In a second aspect, an embodiment of the present invention provides a method for preparing a green pesticide based on glucose lipids and their derivatives. When the water-based pesticide formulation is an emulsion in water, the preparation method comprises:

[0027] Dissolve 10% to 45% of the original pesticide in a solvent until the original pesticide is dissolved; then add 5% to 15% of an emulsifier and stir evenly; after a uniform solution is formed, add 5% to 15% of glucose lipids and their derivatives to disperse them evenly, add 0% to 5% of other surfactants, and continue to stir evenly; then slowly add water to the mixed solution to make up 100%, adjust the pH value to 6-8, and form a uniformly dispersed aqueous emulsion.

[0028] In a third aspect, an embodiment of the present invention provides a method for preparing a green pesticide based on glucose lipids and their derivatives. When the water-based pesticide formulation is a water suspension concentrate, the preparation method comprises:

[0029] Adopt wet grinding ultrafine grinding process, mix 10% to 45% of pesticide technical, 1% to 20% of glucose ester and its derivatives, 4% to 10% of wetting and dispersing aids with water, transfer to grinder for grinding, when more foaming occurs, 0.1% to 0.5% of defoamer can be added, and the grinding medium is glass beads;

[0030] 0.2% to 0.5% of thickener and 1% to 3% of antifreeze are added to the uniformly ground water suspension, and water is supplemented to 100% for grinding to obtain the water suspension.

[0031] In a fourth aspect, an embodiment of the present invention provides an application of a green pesticide based on glucose lipids and their derivatives in the prevention and treatment of agricultural plant diseases including wheat fusarium head blight, wheat root rot, rice sheath blight, and cherry mildew.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] 1. The present invention is based on the application of glucose lipids and their arginine salt derivatives as pesticide adjuvants in pesticides. Compared with common chemical surfactant adjuvants, they exhibit low or non-toxic, biodegradable, environmentally friendly and other characteristics. Compared with common biological surfactants, they have stronger antibacterial activity and good surface / interfacial activity after chemical modification. They have the multifunctional properties of enhanced penetration and antibacterial properties and are used as green and efficient pesticide adjuvants, providing new ideas for the creation of green pesticides for green prevention and control of plant diseases.

[0034] 2. The present invention is based on the application of glucose lipids and their arginine salt derivatives as biological pesticides in pesticides. While having excellent bactericidal, emulsifying, dispersing, penetrating and other properties, it maintains good biosafety, can effectively inhibit a variety of fungal plant diseases, can effectively improve the efficiency of pesticide application, and achieve "reduced application and increased efficiency" of pesticides. It is green, efficient and easily degraded by the environment, thereby reducing the negative impact on the environment.

[0035] 3. The present invention is based on glucose lipids and their derivatives as biological pesticides / adjuvants and compounded with different types and concentrations of pesticide original drugs to prepare a variety of water-based pesticide formulations, such as aqueous emulsions, aqueous suspensions, etc., so that they can play the role of dispersion and penetration promotion while having antibacterial activity. The control effect is 90% to 100%, and the amount applied is 1% to 40% of that of ordinary pesticides. It is green, efficient, degradable, and used in plant disease control. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative labor.

[0037] Figure 1 A schematic flow chart of a method for preparing green pesticides based on glucose lipids and their derivatives provided in an embodiment of the present invention;

[0038] Figure 2 A schematic diagram of the structure of glucose ester (GLs) and its arginine methyl ester derivative (GLs-D-Arg-OMe) provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] The present invention provides a green pesticide based on glucose lipid and its derivatives, comprising:

[0041] One or more of glucose lipids and their derivatives are mixed in any proportion as a first component, and compounded with a second component to obtain a water-based pesticide formulation;

[0042] Wherein, the second component is selected from the pesticide technical including tebuconazole, zinc thiazole, thiophanate-copper, colistin sulfate, and validamycin.

[0043] It should be noted that in this example, the glucose lipid is obtained by chemical modification combined with organic solution extraction from natural crude sophora fermentation biosurfactant, and a chemical modification strategy is further adopted to perform amide condensation modification on the carboxyl terminus of the glucose lipid, and the D-arginine salt group is grafted onto the glucose lipid molecule through a DCC condensation reaction to obtain a glucose lipid D-arginine salt methyl ester derivative. Figure 2 The structure diagram of glucose lipid (GLs) and its arginine salt methyl ester derivative (GLs-D-Arg-OMe) is shown. In this example, glucose lipid and its derivatives are used as biological pesticides for preventing and controlling plant diseases. At the same time, the glucose lipid and its derivatives can be used as pesticide adjuvants and have the functions of emulsification, wetting, dispersion, etc.

[0044] Furthermore, when only glucose lipid is selected as the first component, the mass volume ratio of the glucose lipid in the water-based pesticide formulation is 1 to 40%, preferably 1 to 20%. When only glucose lipid derivative is selected as the first component, the glucose lipid derivative is a glucose lipid D-arginine salt methyl ester derivative, and the mass volume ratio of the glucose lipid derivative in the water-based pesticide formulation is 1 to 80%, preferably 1 to 40%, and more preferably 1 to 20%.

[0045] Furthermore, when the water-based pesticide formulation is an emulsion in water, it includes, by mass volume ratio: 5% to 25% of the original pesticide mass volume ratio, 15% to 40% of the solvent, 5% to 15% of the emulsifier, 5% to 15% of the glucose lipid and its derivatives, 0% to 5% of other surfactants, and the remaining content is supplemented by water to 100%; wherein, the glucose lipid and its derivatives are a mixture of one or more glucose lipids and their arginine salt derivatives in different proportions and concentrations, with a concentration of 1 to 20 g / L.

[0046] Furthermore, the solvent is one or more of xylene, dichloromethane, cyclohexanone, dimethylformamide (DMF), alkylamides, and dimethyl sulfoxide (DMSO) mixed in any proportion. The emulsifier is one or more of phosphate salts, polyoxyethylene ethers, and fatty alcohol polyethers mixed in any proportion.

[0047] Furthermore, when the water-based pesticide formulation is a water suspension, it includes, by mass volume ratio: 10% to 45% of pesticide original, 1% to 20% of glucose lipids and their derivatives, 4% to 10% of wetting and dispersing aids, 0.1% to 0.5% of defoaming agents, 0.2% to 0.5% of thickeners, 1% to 3% of antifreeze agents, and the remaining content is supplemented by water to 100%; wherein, glucose lipids and their derivatives are mixed components of one or more glucose lipids and their arginine salt derivatives in different proportions and concentrations, with a concentration of 1 to 20 g / L.

[0048] Furthermore, the wetting and dispersing aid is one or more of alkylnaphthalene sulfates and lignin sulfonates mixed in any proportion; the defoamer includes silicone defoamer, silicone oil defoamer, silicone defoamer; the thickener is one or more of xanthan gum and magnesium aluminum silicate mixed in any proportion; the antifreeze includes ethylene glycol, glycerin, propylene glycol, and polyvinyl alcohol.

[0049] like Figure 1 As shown, an embodiment of the present invention provides a method for preparing green pesticides based on glucose lipids and their derivatives. When the water-based pesticide formulation is an aqueous emulsion, the preparation method comprises:

[0050] Dissolve 10% to 45% of the pesticide technical in a solvent until the pesticide technical is dissolved; then add 5% to 15% of an emulsifier and stir evenly; after a uniform solution is formed, add 5% to 15% of one or two glucose esters and their derivatives as adjuvants. The glucose esters and their derivatives can be first dissolved in a small amount of water and then slowly added to the pesticide solution while stirring to ensure uniform dispersion. If the emulsification effect is not good, add 0% to 5% of other surfactants and continue to stir evenly; then slowly add water to the mixed solution to make up 100%, place the mixture at room temperature (25-28°C), and adjust the pH value to 6-8 to ensure the stability and safety of the aqueous emulsion, thereby forming a uniformly dispersed aqueous emulsion.

[0051] Furthermore, when the water-based pesticide formulation is an aqueous suspension, the preparation method comprises:

[0052] Adopt wet grinding ultrafine grinding process, mix 10% to 45% of pesticide technical, 1% to 20% of glucose ester and its derivatives, 4% to 10% of wetting and dispersing aids with water, transfer to grinder for grinding, when more foaming occurs, 0.1% to 0.5% of defoamer can be added, and the grinding medium is glass beads;

[0053] 0.2% to 0.5% of thickener is added to the uniformly ground water suspension to adjust the viscosity of the preparation and improve the stability of the preparation, 1% to 3% of antifreeze is added to facilitate its storage and application, and water is supplemented to 100% for grinding to obtain a water suspension.

[0054] Furthermore, the embodiments of the present invention also provide a green pesticide based on glucose lipids and their derivatives for use in preventing and controlling agricultural plant diseases including wheat fusarium head blight, wheat root rot, rice sheath blight, and cherry mildew.

[0055] The experimental process of the present invention is described below:

[0056] 1. Determination of antibacterial test (MIC / MBC): Liquid culture antibacterial test was used to determine the minimum inhibitory concentration of glucose lipids, glucose lipid arginine methyl ester derivatives, and each pesticide original drug using the 96-well plate micro-dilution method. The specific operation is as follows: Use a 2-fold concentration of culture medium (LB culture medium is used for bacteria and PDW culture medium is used for fungi) to dilute the bacterial solution activated for 24-48 hours to an OD of 600 =0.02. Add 100 μL of antibacterial solution and 100 μL of diluted bacterial solution to the 96-well plate and mix well. Finally, the bacterial concentration in the 96-well plate is OD 600 =0.01, the concentration of the antibacterial agent is 0-1024 mg / L. The 96-well plate is fixed in a shaker and cultured under the corresponding conditions, and its OD is measured after 12 hours. 600The inhibition rate (%) of each antibacterial agent on microorganisms was calculated, and its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were summarized.

[0057] 2. Mycelium growth inhibition test: In the clean bench, the sterilized culture medium is made into an antibacterial culture medium plate, and the concentration of the antibacterial agent mother solution is prepared to be 20g / L. After mixing, it is poured into each subpackage culture medium to make the final concentration of the antibacterial agent in each liquid culture medium 0-1024mg / L. The control group is added with an equal amount of pure water. Under sterile conditions, a certain number of bacterial cakes are punched out with a 0.2cm diameter puncher for the test strain, and the bacterial cakes are placed on the antibacterial culture dish with an inoculation needle, with the mycelium side facing down, and each dish is placed in the center, then covered and marked. Each treatment is repeated 3 times, and placed in an inverted culture in a 28℃ constant temperature incubator. After 48 hours, the colony diameter is measured, and the inhibition rate of mycelium growth is recorded and calculated. The colony diameter adopts the cross method to measure the two vertical cross diameters of the colony and calculate their average value.

[0058]

[0059] 3. Spore suspension inhibition experiment: The fungus was evenly spread on the PDA medium, and cultured at a constant temperature of 28°C for 24 hours. An appropriate amount of physiological saline was added to the surface of the medium to wash off the spores. The mycelium was filtered with sterile gauze to obtain a spore suspension. The spore suspension was counted with a hemocytometer. The spore suspension was diluted and adjusted with physiological saline for standby use. The antibacterial agent mother solution was prepared with DMSO to be 20 g / L. 100 μL of the antibacterial agent and 100 μL of 2 times PDW medium were added to a 96-well plate and mixed evenly. The final antibacterial agent concentration of the 96-well plate was 0-1024 mg / L. Pure water and 2.5% DMSO were used as controls. Three parallel experimental and control groups were set up to calculate the minimum inhibitory concentration (MIC) of the fungal spore suspension.

[0060] 4. Control effect experiment: The biological activity of the agent was determined by the spore suspension spray inoculation method. Potted plants (rice and wheat) with the same growth were selected, one leaf was sprayed with the agent and then dried naturally. After 24 hours, the spores of the diseased leaf were washed with distilled water to prepare a spore suspension (10 5 ~10 6 The spore suspension was evenly sprayed on rice / wheat leaves and incubated at room temperature (25-28°C). After 3 days, a graded survey was conducted based on the disease status of the blank control, and the control effect was calculated based on the disease index.

[0061] Grading standard: 0-no disease;

[0062] Level 1: The lesion area accounts for less than 5% of the entire leaf area;

[0063] Level 2: The lesion area accounts for 6-20% of the entire leaf area;

[0064] Level 3: The lesion area accounts for 21-40% of the entire leaf area;

[0065] Level 4: The lesion area accounts for 41-100% of the entire leaf area;

[0066] Disease index = ∑ (number of diseased leaves at each level × relative index) × 100 / (total number of leaves × 4)

[0067]

[0068] The following is a detailed description of the green pesticide based on glucose lipids and their derivatives and the significant effects thereof proposed by the present invention in conjunction with the accompanying drawings. In the absence of conflict, the features of the following embodiments and implementations may be combined with each other.

[0069] Example 1: Preparation of glucose lipids

[0070] Starting from the natural sophorolipid mixture, methanol solution was added to the mixture to prepare a crude sophorolipid methanol solution with a concentration of 50 g / L, and mixed evenly to obtain a clear solution. 3% volume of concentrated sulfuric acid was added to the mixed solution, and the mixed solution was stirred and reacted at 60°C for 10 hours to obtain a glucose lipid methyl esterification product. After the reaction solution was cooled to room temperature, n-hexane and water were added thereto so that n-hexane: water: methanol = 3:2:1, and after mixing evenly, the hydrophilic phase was collected by centrifugation to remove the organic phase.

[0071] Add sodium hydroxide solution to the hydrophilic phase, adjust the pH of the solution to alkaline, stir and react at room temperature for 12 hours to prepare a crude glucose lipid product. Add dilute hydrochloric acid solution to the alkaline reaction solution to adjust the pH value to acidic, and a white precipitate is precipitated, and the precipitate is collected by centrifugation. Add an appropriate amount of pure water to the precipitate for washing, and freeze-dry to obtain a pure glucose lipid product.

[0072] Example 2: Preparation of glucose lipid D-arginine salt methyl ester derivative

[0073] Weigh 1mmol of the prepared protonated glucose lipid, add 20mL of dimethylformamide (DMF) as a solvent, mix well, add 3mmol of dicyclohexylcarbodiimide (DCC) and 1.5mmol of 1-hydroxybenzotriazole (HOBt) to the mixture, and suspend and mix evenly until clear. Add 1.5mmol of arginine methyl ester dihydrochloride and 2mmol of triethylamine, and stir the mixture at room temperature for 24 hours. After the reaction is completed, place the product DMF phase in a refrigerator, filter and remove the precipitated solid until no more precipitation is precipitated in the solution. Add 4-5 times of ethyl acetate to the mixed solution, centrifuge to obtain a crude product of glucose lipid D-arginine salt methyl ester derivative. Add chloroform: isopropanol = 2:1 to the crude product for extraction, place it in a 50°C vacuum drying oven to remove the solvent to obtain a pure product of glucose lipid D-arginine salt methyl ester derivative.

[0074] Example 3: Inhibition of wheat pathogens by glucose lipids and their derivatives

[0075] a) Glucose lipids and their derivatives inhibit mycelium from wheat fusarium rust and root rot

[0076] Material preparation: Glucose lipid (GLs), glucose lipid derivative (GLs-D-Arg-OMe)

[0077] Experimental steps: The wheat pathogens used are wheat fusarium head blight and wheat root rot. Wheat fusarium head blight and root rot are inoculated into potato agar medium (PDA) and cultured for 2-3 days until the mycelium covers the entire medium. Prepare the antibacterial agent PDA medium to make the final concentration of 0-1024 mg / L, let it stand to cool and solidify, and set aside. Inoculate 2 mm wheat fusarium head blight and root rot fungus cakes in the center of the antibacterial medium, culture at 28-30℃ for 48 hours, measure the diameter of each group of colonies, and calculate the inhibition rate.

[0078] The inhibition rate of GLs on the mycelium growth of wheat Gibberella was 80.91% at a concentration of 1024 mg / L. At the same concentration, the inhibition rate of GLs on the mycelium growth of root rot was 77.27%. At the same concentration, the mycelium growth rates of GLs-D-Arg-OMe derivatives on wheat Gibberella and root rot were 98.92% and 96.73%, respectively, showing excellent inhibitory activity.

[0079] Table 1 Inhibition rate of GLs on mycelial growth of wheat pathogens

[0080]

[0081] Table 2 Inhibition rate of mycelial growth of wheat pathogens by GLs-D-Arg-OMe

[0082]

[0083]

[0084] b) Glucose lipids and their derivatives inhibit wheat fusarium spores and root rot

[0085] Material preparation: glucose lipid (GLs), glucose lipid derivative (GLs-D-Arg-OMe); the final concentration range of the antibacterial agent is set to 0-1024 mg / L and placed in a refrigerator for use.

[0086] Spore suspension: After washing the spores of wheat fusarium and root rot with saline, resuspend them in 2 times PDW medium. The number of spores is about 10 6 ~10 7 CFU / mL, for future use.

[0087] Experimental steps: 100 μL of antibacterial agent and 100 μL of 2-fold PDW medium were added to the 96-well plate and mixed evenly. The final antibacterial agent concentration of the 96-well plate was 0-1024 mg / L. Pure water and 2.5% DMSO were used as controls. Three parallel experimental and control groups were set up and placed in a shaker for 4-5 days to observe the growth of the spore suspension. GLs had a weak inhibition on wheat pathogen spores after 5 days. The inhibition rates of wheat fusarium head blight and root rot were 40.91% and 39.73% at the highest concentration, respectively; while the inhibition rate of GLs-D-Arg-OMe on wheat pathogens was greater than 90% at concentrations of 128 mg / L and above, and greater than 99% at concentrations of 256-1024 mg / L. After 5 days of culture, the spore suspension remained clear, showing excellent antibacterial activity, and the minimum inhibitory concentration (MIC) of the spore suspension was 128 mg / L.

[0088] Table 3 Spore inhibition rate of GLs on wheat pathogens

[0089]

[0090] Table 4 Spore inhibition rate of GLs-D-Arg-OMe on wheat pathogens

[0091]

[0092]

[0093] In summary, the glucose lipid derivatives modified by amide condensation reaction have excellent antibacterial properties, can significantly inhibit the growth of wheat pathogens, and can be used as new green pesticides in pesticides.

[0094] Example 4 Glucose lipids and their derivatives compounded with pesticide technical

[0095] Glucose lipids and their derivatives have antibacterial properties superior to general biosurfactants, and have the characteristics of dispersion, wetting, solubilization, etc. to promote the absorption of pesticides by plants. Glucose lipids and their derivatives have significantly improved hydrophilicity and good stability at different temperatures and in different environments. As green and multifunctional biological pesticides, they fully meet the standards of green chemicals and can be widely used in various pesticide preparation scenarios, such as tebuconazole, zinc thiazole, thiophanate-copper, colistin sulfate, and jinggangmycin. The optimal compounding ratio was determined by a 96-well plate antibacterial experiment, and glucose lipids and their derivatives were compounded with pesticides in different ratios. The mass volume ratios of glucose lipids and their derivatives to colistin sulfate and jinggangmycin were set to: 1:100, 1:20, 1:10, 1:5, 1:1, 5:1, 10:1, and 20:1.

[0096] Table 5 Inhibition rate of different proportions of glucose lipids and derivatives combined with pesticide originals on rice pathogens

[0097]

[0098]

[0099] The mass volume ratios of glucose lipids and derivatives to tebuconazole, zinc thiazole, and thiophanate-methyl were set as 1:80, 1:20, 1:10, 1:5, 1:1, 5:1, 10:1, and 20:1.

[0100] Table 6 Inhibition rate of different proportions of glucose lipids and derivatives combined with pesticides on rice pathogens

[0101]

[0102]

[0103] Table 7 Inhibition rate of different proportions of glucose lipids and derivatives combined with pesticides on wheat pathogens

[0104]

[0105] When the combined inhibition rate is higher than 90%, it is judged to have a good pathogenic bacteria joint inhibition effect. When glucose lipid: colistin sulfate = 1:1, the inhibition rate is 90.78%; when glucose lipid: validamycin = 1:1, the inhibition rate is 90.54%; when the ratio of glucose lipid to zinc thiazole, thiophanate-methyl, and tebuconazole is 1:1, the inhibition rates are 92.49%, 90.95%, and 90.75%, respectively. In summary, when the ratio of glucose lipid is 1-50%, there is a good inhibition effect, in which 10% of other additives are added to keep the system stable, so the effective content of glucose lipid is 1-40%.

[0106] When the ratio of glucose lipid derivatives to colistin sulfate and validamycin was 10:1, the inhibition rates were 92.83% and 90.23%; when the ratio of glucose lipids to zinc thiazole, thiophanate-methyl and tebuconazole was 10:1, the inhibition rates were 90.65%, 93.12% and 93.67%. In summary, when the ratio of glucose lipid derivatives was 1-90%, there was a better inhibition effect, and 10% of other additives were added to keep the system stable, so the effective content of glucose lipid derivatives was 1-80%.

[0107] The minimum inhibitory concentration (MIC) of the glucose lipid derivative GLs-D-Arg-OMe against rice pathogens was determined to be 128 mg / L. The specific application cases are as follows:

[0108] A. Colistin sulfate is a polypeptide antibacterial drug with strong antibacterial effect on Gram-negative bacteria. It is often used to inhibit diseases caused by Gram-negative bacteria, but its antibacterial spectrum is narrow and it has no inhibitory effect on Proteus, Brucella, Serratia and all Gram-positive bacteria. According to laboratory tests, at OD 600 =0.01, the minimum inhibitory concentration (MIC) of colistin sulfate against rice pathogens is 32-64 mg / L. Colistin sulfate and GLs-D-Arg-OMe derivatives were mixed in a ratio of 1:1, and the inhibitory effect of glucose lipid derivatives and pesticide raw materials on rice pathogens was detected by turbidimetry. The inhibitory concentration of glucose lipid derivatives was set to 16-128 mg / L,

[0109] The inhibitory concentration of colistin sulfate is 0.5-16 mg / L (the highest inhibitory concentration is 0.5 MIC concentration).

[0110] When the concentration of glucose lipid derivatives is 64 mg / L, only 0.5 mg / L of colistin sulfate is needed to achieve an inhibition rate of more than 98% on rice pathogens. The inhibitory concentration of glucose lipid derivatives is 1 / 2 of the MIC, while the inhibitory concentration of colistin sulfate is only 1 / 128 to 1 / 64 of the MIC, which significantly reduces the amount of pesticide applied and achieves the inhibition of rice pathogens.

[0111] B. Jinggangmycin is a broad-spectrum antibiotic, also known as neomycin. Its chemical structure is a helical polycyclic peptide. It has strong antibacterial activity and can be used to treat infections caused by many bacteria. The metabolites produced by actinomycetes can effectively prevent and control bacterial diseases of rice. According to laboratory tests, at OD 600=0.01, the minimum inhibitory concentration (MIC) of Jinggangmycin against rice pathogens is 5-6g / L. Jinggangmycin and GLs-D-Arg-OMe derivatives were mixed evenly in a ratio of 1:1, and the inhibitory effect of glucose lipid derivatives and pesticide raw materials on rice pathogens was detected by turbidimetry. Jinggangmycin was dissolved in water to prepare an antibacterial agent for use, and the final inhibitory concentration was set to 20-640mg / L, and the inhibitory concentration of glucose lipid derivatives was 16-128mg / L (both less than the MIC value).

[0112] When the concentration of glucose lipid derivatives is 64 mg / L, only 20 mg / L of jinggangmycin is needed to achieve an inhibition rate of more than 99% on rice pathogens. The inhibition concentration of glucose lipid derivatives is 1 / 2 of the MIC, while the inhibition concentration of jinggangmycin is only 1 / 300 to 1 / 250 of the MIC concentration compared with the MIC. Compared with the inhibition of rice pathogens by pesticides alone, the amount of pesticide required for the inhibition concentration is significantly reduced after the pesticide is compounded with the glucose lipid arginine salt derivative. At the same time, as a biosurfactant, it is safe, green and easily degraded by the environment. It can be used in the field of green pesticides, effectively reducing the application amount of pesticides, achieving the "reduced application and increased efficiency" of pesticides and reducing the negative impact on the environment.

[0113] Example 5 Stability of glucose lipids and their derivatives combined with pesticide originals

[0114] The pesticide technical was dissolved in an organic solvent, and the glucose ester and its derivatives were dissolved in water, wherein the concentration of the glucose ester and its derivatives mother liquor was 20 g / L. When the volume ratio of the organic phase: the aqueous phase was set to 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1, a certain amount of emulsifier or wetting dispersant was added to make it stable, and the stability of the preparation was tested under room temperature, cold storage and hot storage environments.

[0115] Among them, the mass volume ratio of the organic phase (containing the original pesticide) is 5g / L, and the mass volume ratio of the aqueous phase (divided into glucose lipids, glucose lipid derivatives, and a mixture of glucose lipids and their derivatives) is 5g / L. The stability of the system is shown in the following table. When the aqueous phase ratio of glucose lipids and their derivatives is 1-40%, the system remains stable; when the aqueous phase ratio of glucose lipids and their derivatives is 1-20%, the system remains stable for 48 hours, so it is further preferably 1-20%.

[0116]

[0117] Table 8 Determination of aqueous stability of glucose lipids and derivatives

[0118] Example 6 Glucose lipids and their derivatives used as green pesticide formulations

[0119] Glucose lipids and their derivatives have antibacterial activity superior to that of ordinary biosurfactants, and also have the properties of dispersion, wetting, solubilization, and emulsification. They can completely replace chemical surfactants and be used in pesticides as multifunctional green pesticides.

[0120] Various water-based pesticide formulations, such as aqueous emulsions and aqueous suspensions, can be prepared by compounding glucose lipids and their derivatives with different types and concentrations of pesticide technicals. Specific examples are as follows:

[0121] 1) Preparation of Tebuconazole-Glucose Lipid Emulsion in Water

[0122] Tebuconazole belongs to the triazole fungicide class and is a thiol demethylation inhibitor. It can effectively prevent and treat various rusts, powdery mildew, web blotch, root rot, and fusarium head blight of cereal crops. However, the current pesticide formulations require more organic solvents. Dissolve tebuconazole in a mixed solvent of xylene, cyclohexanone, and DMF until the pesticide is dissolved. Add 8% of emulsifier and stir evenly. After forming a uniform solution, add glucose ester aqueous solution (2g / L) slowly into the pesticide solution and continue to stir evenly. Then slowly add water to the mixed solution to make up 100%, forming a uniform, milky 20% tebuconazole glucose ester aqueous solution. Its appearance is a milky white solution, which remains stable at different temperatures (-20℃-40℃) and different pH values ​​(6-8). Compared with the same concentration of tebuconazole pesticide on the market, the application amount is only 10% to 40% to achieve similar effects. The prevention and control effect on plant diseases such as wheat pathogens is more than 95%, which can effectively improve the application efficiency of pesticides and reduce the negative impact on the environment.

[0123] The specific formula is shown in the table below. The formula is left standing at room temperature for 24 hours, and the demulsification rate is less than 3%.

[0124]

[0125] 2) Preparation of colistin sulfate·GLs-D-Arg-OMe aqueous suspension

[0126] Colistin sulfate can effectively inhibit Gram-negative bacteria and has a strong inhibitory effect on rice pathogens, but the amount of application currently sold on the market is relatively high. Glucose lipid D-arginine salt methyl ester derivative (GLs-D-Arg-OMe) has excellent antibacterial activity. Using a wet grinding ultrafine grinding process, colistin sulfate, GLs-D-Arg-OMe, a wetting dispersant (B) and water are mixed, transferred to a grinder and ground for 2-3 hours, and 0.2% defoaming agent is added until the grinding is uniform. Adding 0.5% xanthan gum improves the stability of the preparation; adding 1% antifreeze helps its storage and application, and the water is supplemented to 100% for research and development to obtain colistin sulfate·GLs-D-Arg-OMe water suspension. The suspension did not show bottom precipitation within 1 hour, and remained stable at different temperatures and pH values. Compared with the water suspensions of the same concentration on the market,

[0127] The application amount is only 5% to 10% of that of the above compound to achieve similar effects, and the control effect is 100%. The specific formula is as follows:

[0128]

[0129]

[0130] *The preparation process of Jinggangmycin·GLs-D-Arg-OMe aqueous suspension is the same as above

[0131] 3) Preparation of aqueous suspension of zinc thiazole·glucose ester and its derivatives

[0132] Thiazole zinc has both fungicidal and bactericidal effects, good control effects, and a wide range of control targets, but the current required pesticide application amount is relatively high. Thiazole zinc, GLs-D-Arg-OMe, glucose ester and water are mixed by wet grinding and ultrafine grinding technology to obtain a water suspension concentrate of thiazole zinc glucose ester and its derivatives. The suspension concentrate does not show bottom precipitation within 1 hour and remains stable at different temperatures and pH values. Compared with the water suspension concentrate of the same concentration on the market, the application amount is only 1% to 10% to achieve similar effects, and the control effect on plant pathogens is 100%. The specific formula is as follows:

[0133]

[0134] *The preparation process of thiophanate-methyl·glucose ester and its derivatives aqueous suspension is the same as above

[0135] Matters not covered by the present invention are known technologies.

[0136] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology to understand the content of the present invention and implement it accordingly. They are only preferred examples of the present invention and cannot be used to limit the protection scope of the present invention. In addition to the above embodiments, the present invention can also have other implementation methods. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A green pesticide based on glucose lipids and their derivatives, characterized in that: include: One or more of glucose lipids and their derivatives are mixed in any proportion as a first component, and compounded with a second component to obtain a water-based pesticide formulation; Wherein, the second component is selected from the pesticide technical including tebuconazole, zinc thiazole, thiophanate-copper, colistin sulfate, and validamycin.

2. A green pesticide based on glucose lipids and their derivatives according to claim 1, characterized in that: When only glucose lipid is selected as the first component, the mass volume ratio of the glucose lipid in the water-based pesticide formulation is 1-40%.

3. A green pesticide based on glucose lipids and their derivatives according to claim 1, characterized in that: When only glucose lipid derivatives are selected as the first component, the glucose lipid derivatives are glucose lipid D-arginine salt methyl ester derivatives, and the mass volume ratio of the glucose lipid derivatives in the water-based pesticide formulation is 1 to 80%.

4. A green pesticide based on glucose lipids and their derivatives according to claim 1, characterized in that: When the water-based pesticide formulation is an emulsion in water, it shall include, by mass volume ratio: The mass volume ratio of pesticide technical is 5% to 25%, solvent is 15% to 40%, emulsifier is 5% to 15%, glucose lipid and its derivatives are 5% to 15%, other surfactants are 0% to 5%, and the rest is made up to 100% by water; Wherein, the concentration of glucose lipid and its derivatives is 1-20g / L.

5. A green pesticide based on glucose lipids and their derivatives according to claim 4, characterized in that: The solvent is one or more of xylene, dichloromethane, cyclohexanone, dimethylformamide, alkylamides, and dimethyl sulfoxide mixed in any proportion; The emulsifier is one or more of phosphate salts, polyoxyethylene ethers, and fatty alcohol polyethers mixed in any proportion.

6. A green pesticide based on glucose lipids and their derivatives according to claim 1, characterized in that: When the water-based pesticide formulation is a water suspension concentrate, it shall include, by mass volume ratio: Pesticide technical 10% to 45%, glucose ester and its derivatives 1% to 20%, wetting and dispersing aid 4% to 10%, defoamer 0.1% to 0.5%, thickener 0.2% to 0.5%, antifreeze 1% to 3%, and the rest is made up to 100% by water; Wherein, the concentration of glucose lipid and its derivatives is 1-20g / L.

7. A green pesticide based on glucose lipids and their derivatives according to claim 6, characterized in that: The wetting and dispersing aid is one or more of alkylnaphthalene sulfates and lignin sulfonates mixed in any proportion; The defoaming agent includes organic silicon defoaming agent, silicone oil defoaming agent, silicone defoaming agent; The thickener is one or more of xanthan gum and magnesium aluminum silicate mixed in any proportion; The antifreeze agent includes ethylene glycol, glycerol, propylene glycol, and polyvinyl alcohol.

8. A method for preparing green pesticides based on glucose lipids and their derivatives, characterized in that: When the water-based pesticide formulation is an emulsion in water, the preparation method comprises: Dissolve 10% to 45% of the original pesticide in a solvent until the original pesticide is dissolved; then add 5% to 15% of an emulsifier and stir evenly; after a uniform solution is formed, add 5% to 15% of glucose lipids and their derivatives to disperse them evenly, add 0% to 5% of other surfactants, and continue to stir evenly; then slowly add water to the mixed solution to make up 100%, adjust the pH value to 6-8, and form a uniformly dispersed aqueous emulsion.

9. A method for preparing green pesticides based on glucose lipids and their derivatives, characterized in that: When the water-based pesticide formulation is an aqueous suspension, the preparation method comprises: Adopt wet grinding ultrafine grinding process, mix 10% to 45% of pesticide technical, 1% to 20% of glucose ester and its derivatives, 4% to 10% of wetting and dispersing aids with water, transfer to grinder for grinding, when more foaming occurs, 0.1% to 0.5% of defoamer can be added, and the grinding medium is glass beads; 0.2% to 0.5% of thickener and 1% to 3% of antifreeze are added to the uniformly ground water suspension, and water is supplemented to 100% for grinding to obtain the water suspension.

10. Application of green pesticides based on glucose lipids and their derivatives in the prevention and treatment of agricultural plant diseases including wheat fusarium head blight, wheat root rot, rice sheath blight and cherry mildew.

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