阅读说明：本技术 一种海藻酸寡糖与春雷霉素的组合杀菌剂及应用 (Composite bactericide of alginate oligosaccharide and kasugamycin and application ) 是由 尹恒 刘启顺 于 2019-11-08 设计创作，主要内容包括：本发明公开了一种含有海藻酸寡糖和春雷霉素的组合杀菌物,具体涉及一种农药组合物,它以海藻酸寡糖和春雷霉素组合作为活性成分。其包含下述重量百分比有效成分的组合物海藻酸寡糖为与春雷霉素的质量比为0.5-15：0.1-30,优选2-10：5-20,助剂余量。该组合杀菌剂可广泛用于防治蔬菜、水稻、果树等病菌的危害,对作物具有保护和治疗作用,表现出极强的抗菌活性。试验证明本发明所提供的杀菌组合物既具有诱抗活性、又具有杀菌活性,具有杀菌谱广、使用成本低、对作物及环境使用安全等特点。(The invention discloses a composite bactericide containing alginic acid oligosaccharide and kasugamycin, and particularly relates to a pesticide composition which takes the combination of alginic acid oligosaccharide and kasugamycin as active ingredients. The composition of alginic acid oligosaccharide and kasugamycin comprises the following effective components in percentage by weight, wherein the mass ratio of the alginic acid oligosaccharide to the kasugamycin is 0.5-15: 0.1 to 30, preferably 2 to 10: 5-20 percent of additive and the balance of auxiliary agent. The composite bactericide can be widely used for preventing and treating the harm of germs of vegetables, rice, fruit trees and the like, has the functions of protecting and treating crops, and shows strong antibacterial activity. Experiments prove that the bactericidal composition provided by the invention has the advantages of inductive and anti-activity, bactericidal activity, broad bactericidal spectrum, low use cost, safe use for crops and environment and the like.)

1. A composite bactericide of alginate oligosaccharide and kasugamycin is characterized in that: the composition contains active ingredients of alginic acid oligosaccharide and kasugamycin, wherein the mass ratio of the alginic acid oligosaccharide to the kasugamycin is (0.5-15): 0.1 to 30, preferably 2 to 10: 5-20.

2. The composite germicide according to claim 1, wherein: the alginic acid oligosaccharide is a sodium alginate degradation product, is a copolymer which is formed by connecting alpha-L-mannuronic acid (M) and beta-D-guluronic acid (G) through 1, 4-glycosidic bonds and consists of different G and M fragments, and has a molecular formula of (C)₆H₇O₆Na) n, and the polymerization degree n is 2-10.

3. The germicidal composition as claimed in claim 1 or 2, wherein: the composite bactericide takes alginic acid oligosaccharide and kasugamycin as main active ingredients and is prepared into any bactericide formulation allowed in pesticides with one or more than two of pesticide auxiliary agents, fillers or solvents;

in the combined bactericide: the mass content of the alginic acid oligosaccharide is 0.5-15%, and the preferred mass content of the alginic acid oligosaccharide is 2-10%; the mass content of kasugamycin is 0.1-30%, and the mass content of kasugamycin is preferably 5-20%.

4. The germicidal composition of claim 3, wherein: the dosage form of the composite bactericide of the alginic acid oligosaccharide and the kasugamycin is preferably one of a soluble solution, an aqueous emulsion, a microemulsion, a suspending agent, a water dispersible granule and a wettable powder.

5. Use of a fungicidal composition according to any of claims 1 to 4 for the control of fungal and/or bacterial diseases of plants.

6. Use according to claim 5, characterized in that: the composite bactericide can prevent and treat one or more than two of bacterial angular leaf spot, rice blast, kiwifruit canker, tomato late blight and gray mold.

7. Use according to claim 5, characterized in that: the combined bactericide can be used for preventing and treating fungal and/or bacterial diseases of rice, vegetables or fruit trees.

8. Use according to any one of claims 5 to 7, characterized in that: the alginate oligosaccharide of claim 3 or 4 is diluted 500-fold and applied 2000-fold in combination with kasugamycin.

9. Use according to any one of claims 5 to 8, characterized in that: the dosage of any one of the combination bactericides is 10-80g of active ingredient per hectare.

Technical Field

The invention belongs to the technical field of pesticide compounding, relates to a composite bactericide, and particularly relates to a composite bactericide containing alginate oligosaccharides and kasugamycin.

Background

The alginic acid oligosaccharide is obtained by enzymolysis of sodium alginate, is composed of alpha-L-mannuronic acid (M) and beta-D-guluronic acid (G) connected by 1, 4-glycosidic bond, and has a molecular formula of (C)₆H₇O₆Na) n, and the polymerization degree n is 2-10. The alginate oligosaccharide has the functions of inducing plant resistance, resisting drought and cold, promoting plant growth, protecting flower and fruit, etcVarious plant physiological activities. Has the advantages of small relative molecular mass, low toxicity, good water solubility, strong absorbability and the like.

Kasugamycin (Kasugamycin), also known as Kasugamycin, has a chemical structure of 5-amino-2-methyl-6- (2, 3, 4, 5, 6-carbonyl cyclohexyl oxo) pyran-3-ylamino-alpha-iminoacetic acid, and belongs to a low-toxicity bactericide of antibiotics. The action mechanism is that an esterase system which interferes with the amino acid metabolism of pathogenic bacteria destroys the biosynthesis of protein, inhibits the growth of hypha and causes cell granulation, and leads the pathogenic bacteria to lose the reproductive and infection capacity, thereby achieving the aim of killing the pathogenic bacteria and preventing and treating diseases. Has activity to bacteria, yeast and plant fungi, has stronger imbibition property, and is an ideal medicament for preventing and treating various bacteria and fungal diseases of crops such as vegetables, melons and fruits.

The bactericide is singly used for a long time, so that the drug resistance of germs is easily generated, the drug effect is obviously reduced, the dosage is greatly increased, the spraying period is shortened, the using effect is poor, and the cost is increased. In addition, the use of a large amount of pesticides makes the resistance of germs worse, causes certain influence on the environment, increases the drug residue of agricultural products and influences the food safety.

The alginate oligosaccharide is a plant resistance inducer and can induce plants to generate disease resistance. Kasugamycin is an esterase system which interferes with the amino acid metabolism of pathogenic bacteria and is a high-efficiency bactericide which destroys the biosynthesis of proteins. The action mechanisms of the two are different, the two are scientifically combined to prepare the combined bactericide, and the combined bactericide can simultaneously have an internal and external synergistic sterilization effect of inducing the enhancement of the internal immunity of plants and simultaneously having a high-activity sterilization effect on the external. The combined bactericide enlarges the bactericidal spectrum of the product, has wide application range, has a synergistic effect on the compounded composition, has good control effect, reduces the dosage and the dosage cost, reduces the adverse effect on the environment, and greatly delays the generation of the drug resistance of germs.

Four patents report the compound combined preparation of amino-oligosaccharin (chitosan oligosaccharide) and kasugamycin (application number: 200710085999.4; application number: 201110125705.2; application number: 201710193393.6; application number: 201811290248), mainly the combination, powder, aqua and application of amino-oligosaccharin and kasugamycin. The amino-oligosaccharin (chitosan oligosaccharide) is basic oligosaccharide with amino, has positive charge, is easy to precipitate with an anionic auxiliary agent or a surfactant in a conventional pesticide preparation, and causes the preparation to be unstable; in addition, in the using process of farmers, when different pesticides are mixed, the compound combined preparation of the amino-oligosaccharin (chitosan oligosaccharide) and the kasugamycin is easy to precipitate with other pesticides, so that the conditions of complex operation, drug loss and the like are caused. More importantly, each oligosaccharide has its own range of elicitation and relative activity.

The alginic acid oligosaccharide is anionic oligosaccharide, and has good compounding property with other pesticides; the composite bactericide of alginic acid oligosaccharide and kasugamycin has better drug effect than the compound composite preparation of amino-oligosaccharide (chitosan oligosaccharide) and kasugamycin in some plant diseases. The composite bactericide of alginic acid oligosaccharide and kasugamycin is not reported in documents and patents.

In order to achieve the above purpose, the following technical scheme is adopted in the patent:

the composite bactericide of alginic acid oligosaccharide and kasugamycin comprises the active ingredients of alginic acid oligosaccharide and kasugamycin, wherein the mass ratio of alginic acid oligosaccharide to kasugamycin is (0.5-15): 0.1 to 30, preferably 2 to 10: 5-20. The alginic acid oligosaccharide is a sodium alginate degradation product, is a copolymer which is formed by connecting alpha-L-mannuronic acid (M) and beta-D-guluronic acid (G) through 1, 4-glycosidic bonds and consists of different G and M fragments, and has a molecular formula of (C)₆H₇O₆Na) n, and the polymerization degree n is 2-10. The composite bactericide takes alginic acid oligosaccharide and kasugamycin as main active ingredients and is prepared into any bactericide formulation allowed in pesticides with one or more than two of pesticide auxiliary agents, fillers or solvents; in the combined bactericide: the mass content of the alginic acid oligosaccharide is 0.5-15%, and the preferred mass content of the alginic acid oligosaccharide is 2-10%; the mass content of kasugamycin is 0.1-30%, and the mass content of kasugamycin is 5-20%. The dosage form of the combined bactericide of the alginate oligosaccharide and the kasugamycin is preferably soluble solution, aqueous emulsion or microemulsionOne of emulsion, suspending agent, water dispersible granule and wettable powder. The bactericidal composition can be used for controlling fungal and/or bacterial diseases of plants. The composite bactericide can prevent and treat one or more than two of bacterial angular leaf spot, rice blast, kiwifruit canker, tomato late blight and gray mold. The combined bactericide can be used for preventing and treating fungal and/or bacterial diseases of rice, vegetables or fruit trees. The alginate oligosaccharide and kasugamycin combined bactericide is diluted by 500-fold and applied by 2000-fold. The dosage of the combined bactericide is 10-80g of active ingredients per hectare.

The pesticide auxiliary agent is one or more than two of alkyl naphthalene formaldehyde condensate sulfonate, nonylphenol polyoxyethylene ether, sodium lignin sulfonate, xanthan gum, an organosilane defoaming agent, PO-EO block polyether, sodium dodecyl sulfate, sodium methoxy fatty acid aminobenzenesulfonate, sodium methylene dinaphthalene sulfonate, TX-IO and AEO-5;

the filler is one or more than two of ammonium sulfate, urea, clay, white carbon black and kaolin;

the solvent is one or more than two of water, dimethyl sulfoxide and N, N-dimethylformamide;

this patent has the advantage:

1. the combination germicide of this patent have the inside immunity of induced plant reinforcing simultaneously, the outside has the inside and outside effect of disinfecting of sterilizing effect of high activity again simultaneously, "sterilizing effect is more high-efficient in coordination.

2. The composite bactericide expands the bactericidal spectrum of products, so that the application range is wide, and the compounded composition has a synergistic effect.

3. The composite bactericide contains the green plant resistance inducer alginate oligosaccharide, reduces the dosage and the medication cost, reduces the adverse effect on the environment, greatly delays the generation of drug resistance of germs, and has high economic and social benefits.

4. The combination bactericide has good mixing performance with other chemical pesticides.

Detailed Description

The present invention is further illustrated at ー in the following examples, which are not intended to be limiting and the dosage forms of the examples are prepared by conventional methods.

Example 1: preparation of 10% alginic acid oligosaccharide and kasugamycin suspending agent

The kasugamycin suspension comprises the following components in parts by weight: 5 percent of alginic acid oligosaccharide (the polymerization degree n is 2-10), 5 percent of kasugamycin, 1 percent of 2-naphthalenesulfonic acid formaldehyde polymer sodium salt, 1 percent of nonylphenol polyoxyethylene ether, 0.3 percent of xanthan gum, 0.3 percent of methyl ethoxy silicone oil defoamer and the balance of water. Mixing all the materials, stirring uniformly, adding the materials into a sand mill, and sanding to ensure that the materials reach the fineness of 2-5 mu m, thus obtaining the 10% alginic acid oligosaccharide-kasugamycin suspending agent.

Example 2: preparation of 15% alginic acid oligosaccharide-kasugamycin water dispersible granules

The kasugamycin water dispersible granule comprises the following components in parts by weight: 2% of alginate oligosaccharide (the polymerization degree n is 2-10), 13% of kasugamycin, 8% of sodium lignin cross, 3% of surfactant polyoxyethylene-polyoxypropylene block polyether, 4% of sodium dodecyl sulfate, 5% of ammonium sulfate, 12% of urea and the balance of clay. The components are uniformly mixed according to the proportion of the formula, are crushed into wettable powder by air flow, are added with a certain amount of water to be mixed, extruded and granulated, and are dried and screened to obtain the 15 percent alginic acid oligosaccharide and kasugamycin water dispersible granule.

Example 3: preparation of 18% alginic acid oligosaccharide and kasugamycin soluble solution

The kasugamycin soluble solution comprises the following components in parts by weight: 10% of alginic acid oligosaccharide (the polymerization degree n is 2-10), 8% of kasugamycin, 1% of nonylphenol polyoxyethylene ether emulsifier (TX-10), 2% of fatty alcohol polyoxyethylene ether (AEO-5), 3% of urea and the balance of water. Mixing and dissolving the components, and then dissolving the components in 18 percent of alginic acid oligosaccharide and kasugamycin soluble solution.

Example 4: preparation of 25% alginic acid oligosaccharide and kasugamycin wettable powder

The kasugamycin wettable powder comprises the following components in parts by weight: 5% of alginic acid oligosaccharide (the polymerization degree n is 2-10), 20% of kasugamycin, 3% of p-methoxy fatty acid sodium aminobenzenesulfonate, 4% of methylene dinaphthalene sodium sulfonate, 5% of white carbon black and the balance of kaolin. And (3) performing jet milling on the components to prepare 25% alginic acid oligosaccharide-kasugamycin wettable powder.

Example 5: drug effect test of combined bactericide on bacterial angular leaf spot of watermelon

The 4 mixed agents obtained in the embodiments 1-4 of the invention are subjected to a pharmacodynamic test, and kasugamycin, alginic acid oligosaccharide and 5% amino-oligosaccharin and 10% kasugamycin water are adopted as positive control, and clear water is adopted as negative Control (CK)

Test materials

The test field is arranged in the three-joint watermelon land in the north sea of Guangxi, the area is about 5 hectares, and the watermelon variety is farmer-friend black beauty. Transplanting 2-leaf 1-heart grafted seedlings in 2018, 7 months and 25 days, and transplanting about 510 plants per mu. The seedlings have 7-10 leaves in the 8-month and 2-day efficacy test.

Design of experiments

There are 8 treatments in total: the concentrations of the active ingredients for the specific applications are shown in table 1. Cell area 30m²3 times of repetition, random block design, total area 720m². Spraying 50kg of the pesticide per mu by using a knapsack sprayer for 1 time in 8 months and 8 days, and uniformly spraying the pesticide on the front and back surfaces of the leaves.

Test investigation and statistical method

And (3) examining disease indexes before pesticide application, randomly extracting 10 watermelons in each cell, fixedly extracting 6 leaves on the main vines of each watermelon for examination, marking, examining before pesticide application each time, grading the area of lesion spots on each leaf in percentage of the whole leaf area, and recording the total leaf number, the leaf number of each grade of disease and the disease grade number. The classification standard is as follows: level 0: no disease spots on the leaves; level 1: the lesion area accounts for less than 5% of the whole leaf area; and 3, level: the lesion area accounts for 6 to 10 percent of the whole leaf area; and 5, stage: the lesion area accounts for 11 to 25 percent of the whole leaf area; and 7, stage: the lesion area accounts for 26-50% of the whole leaf area; and 9, the disease spot area accounts for more than 51 percent of the whole leaf area (the classification standard refers to the pesticide field efficacy test criterion II GB/T17980, 112-plus 2004, the disease is classified according to the severity of the disease, each grade is represented by simple numerical values according to the severity sequence, and then the disease condition is calculated by the following formula according to the recorded disease condition:

disease index ═ Σ (number of diseased leaves at each stage × relative stage)/(total investigated leaves number × 9) × 100%

The control effect is (1-CK pre-drug control disease index multiplied by drug post-treatment disease index in a drug treatment area/(CK post-drug control disease index multiplied by drug pre-treatment disease index in a drug treatment area)) × 100%

TABLE 1 prevention and treatment of bacterial angular leaf spot of watermelon

The test results in table 1 show that the control effect of the alginate oligosaccharide and kasugamycin composition on bacterial angular leaf spot of watermelons is over 80 percent, and is obviously higher than that of a single-dose control medicament and a positive control medicament. Adverse reactions such as phytotoxicity and the like do not occur in each treatment in the test. Therefore, the practical prevention and treatment effect of the combination of the alginic acid oligosaccharide and the kasugamycin is obviously better than that of a single preparation, and the combination of the alginic acid oligosaccharide and the kasugamycin has obvious synergistic effect. Meanwhile, the pesticide effect of the alginic acid oligosaccharide and kasugamycin combined bactericide on bacterial angular leaf spot of watermelons is better than that of the amino-oligosaccharide and kasugamycin combined bactericide.

Example 6: drug effect test of combined bactericide on kiwifruit canker

In 2018, the samples of examples 1-4 were tested in Shaanxi West to county for field control, and kasugamycin, alginate oligosaccharide, 5% amino-oligosaccharide, 10% aqueous solution of kasugamycin and 72% streptomycin sulfate soluble powder were used as positive controls. And selecting disease spots with uniform sizes on the diseased branches as treatment objects, and uniformly distributing each treated disease spot on each branch of the same kiwi fruit tree. The test was carried out with 8 treatments, the specific active ingredient concentrations applied being shown in table 2.

Scraping the epidermis of the selected scab with a knife, measuring the area of the scab, spraying the medicament on the scab with a spray can for 2 times, and continuously spraying 50kg of medicament per mu each time. After 7 days, the examination was carried out, and the lesion surface was measured. The test was performed in 12 lesions per treatment, 3 replicates were set. The relative control effect calculation method is as follows:

TABLE 2 relative control effect of different formulation compositions on kiwifruit canker

Reagent for testing	Final application concentration of active ingredient (. mu.g/mL)	Relative control effect (%)
			Example 1 product	70	85.3
EXAMPLE 2 product	70	90.3
			EXAMPLE 3 product	70	89.7
EXAMPLE 4 product	70	91.2
			10% kasugamycin water aqua	70	73.3
5% alginic acid oligosaccharide aqueous solution	10	60.3
			5% amino-oligosaccharin + 10% kasugamycin (w/w)	70	82.5
72% streptomycin sulfate soluble powder	300	71.5

The test results in table 2 show that the control effect of the alginate oligosaccharide and kasugamycin composition on kiwifruit canker is more than 85 percent and can exceed 90 percent, and the control effect is obviously higher than that of a single-dose control medicament and a positive control medicament. Adverse reactions such as phytotoxicity and the like do not occur in each treatment in the test. Therefore, the practical prevention and treatment effect of the combination of the alginic acid oligosaccharide and the kasugamycin is obviously better than that of a single preparation, and the combination of the alginic acid oligosaccharide and the kasugamycin has obvious synergistic effect. The pesticide effect of the alginic acid oligosaccharide and kasugamycin combined bactericide on kiwifruit canker is better than that of the amino-oligosaccharide and kasugamycin combined bactericide.

Example 7: drug effect test of combined bactericide on rice blast

In 2018, in 5-9 months, the combined bactericide of the embodiments 1-4 of the invention is used for field test of rice blast in Linxiang county in Yueyang city of Hunan province, and kasugamycin, alginic acid oligosaccharide, 5% amino-oligosaccharide and 10% kasugamycin aqua are used as positive controls. The total number of 7 treatments is set, each treatment is repeated for 3 times, and the cell area is 30m²Random block permutation. The concentrations of the active ingredients for the specific applications are shown in table 3. Spraying at early stage of disease onset by conventional spraying methodThe first time of pesticide application, the second time of pesticide application for 1 time at intervals of 7d, the total time of pesticide application is 3 times, and 60kg of pesticide liquid is sprayed each time per mu. The effect of the drug was investigated 14d after the 3 rd dose. Sampling is carried out at 5 points per cell, 50 plants are taken at each point, and each plant investigates the flag leaf and two leaves below the flag leaf.

The percentage of lesion area per leaf or leaf sheath on the whole rice plant was investigated and classified.

Grading standard:

level 0; no disease spots; level 1: 1-5 leaf spots, wherein the length of the spots is less than 1 cm; and 2, stage: 6-10 leaf spots, wherein the length of part of the spots is more than 1 cm; and 3, level: 11-25 leaf spots, wherein part of the spots are connected into pieces, and the area of the spots accounts for 10-25% of the area of the leaves; 4, level: the number of the disease spots on the leaves is more than 26, the disease spots are connected into a tablet, and the area of the disease spots accounts for 26-50% of the area of the leaves; and 5, the disease spots are connected into tablets, and the area of the disease spots accounts for more than 50 percent of the area of the leaves.

Calculating disease index and preventing and treating effect.

Disease index ∑ (leaf incidence rate at each stage X this stage representative value)/(survey total leaf number X highest stage representative value) X100%

The prevention and treatment effect (%) is (1- (pre-drug control disease index X post-drug treatment disease index)/(post-drug control disease index X pre-drug treatment disease index)) X100%

Table 3. controlling effect of each treatment agent against Pyricularia oryzae by 14 days (%)

The field efficacy test result shows that the alginic acid oligosaccharide, kasugamycin and the composition have excellent control effect on rice blast, have synergistic effect compared with a single agent in control effect, and can reduce the use amount of pesticide and reduce the cost. The pesticide effect of the alginic acid oligosaccharide and kasugamycin combined bactericide on rice blast is better than that of the amino-oligosaccharide and kasugamycin combined bactericide.

Example 8: drug effect test of combined bactericide on tomato late blight and gray mold

The combination preparations of examples 1 to 4 as well as the single agents were formulated to the desired concentrations. Before testingCorresponding single agents and blank controls are arranged in the tomato field, the concentration of the active effective components is shown in table 4, and 60kg of liquid medicine is sprayed each time per mu. At the early stage of disease occurrence, the traditional constant spraying of a handheld sprayer is adopted, and 50 plants are sprayed per agent. The application is carried out for 1 time 7-10 days after the application of the first time. Disease conditions of the branded tomato plants are investigated 10-20 days after the application for the 2 nd time. Each plant investigates all the compound leaves, and the disease grade is divided according to the percentage of the lesion area on each compound leaf to the whole compound leaf area. Level 0: no disease spots; level 1: the lesion spots account for less than 5% of the whole compound leaf area; and 3, level: the lesions occupying the whole area of the compound leavesAnd 5, stage: the lesions occupying the whole area of the compound leaves And 7, stage: the lesions occupying the whole area of the compound leavesAnd 9, stage: the lesion spots account for more than 51 percent of the whole compound leaf area.

Disease index ═ Σ (number of diseased leaves at each level X relative to disease level)/(total investigated leaves X9) X100%

The preventing and treating effect (%) is [ (disease index of control area-disease index of treatment area)/disease index of control area ] X100%

TABLE 4 field Effect of kasugamycin-containing fungicidal compositions on tomato diseases

As shown in Table 4, the drug effect of the composition of alginic acid oligosaccharide and kasugamycin on tomato late blight and gray mold is higher than that of a corresponding single dose, and the composition has an obvious synergistic effect. Meanwhile, the pesticide effect of the alginic acid oligosaccharide and kasugamycin combined bactericide on tomato late blight and gray mold is better than that of the amino-oligosaccharide and kasugamycin combined bactericide.