KR20220149045A - An use of bactericidal composition - Google Patents

Abstract

The present invention relates to the use of a bactericidal composition wherein the active ingredients are oxathiapiprolin and cyanoestrobin, a weight ratio of the oxathiapiprolin and cyanoestrobin is 5 : 1 to 1 : 1. The bactericidal composition is effective in preventing and treating late blight of potatoes and has excellent application value.

Description

AN USE OF BACTERICIDAL COMPOSITION

The present invention relates to an agricultural bactericidal composition, in particular to a bactericidal composition comprising oxathiapiprolin and cyanoestrobin as active ingredients and uses thereof. The present invention also relates to the use of the bactericidal composition for the prevention and control of crop diseases, in particular against grape downy mildew and potato late blight, and belongs to the field of pesticides.

Oxathiapiprolin has a unique target site and novel mechanism of action. It is fast-acting, has good durability, and is resistant to rainwater. It can show good control effect even at very low doses. It is very effective against plant diseases caused by oomycete pathogens, for example potatoes, grapes, vegetables and other special crops such as downy mildew of grapes, melons and cabbage, late blight of tomatoes and potatoes, pepper blight and other diseases. effectively control oomycete disease. However, because oxathiapiproline has a single site of action and carries a risk of moderate to high resistance, resistance management is required. Currently, to effectively delay the development of drug resistance and extend the life cycle of drugs, researchers combine or alternate use of oxathiapiproline with fungicides with different mechanisms of action. For example, oxathiapiproline may contain methamines such as azoxystrobin, cresoxim-methyl, pyraclostrobin, thiabendazole, and thiophene ketone. It has been reported to be formulated and used with methoxyacrylate fungicides, but the effect is not significant.

Cyanoestrobin is effective against diseases caused by Fusarium and has protective and therapeutic effects. It is absorbed through the root and has an upward conductivity from the leaf, and the conductivity between the lower part of the leaf and between the leaves is poor. Cyanoestrobin can effectively control stem basal rot, root rot and pith necrosis of tomatoes, grapes and other crops. However, there are few prior art on the control effect on grape downy mildew and potato late blight.

Surprisingly, the applicants have found that the combination of oxathiapiproline, which has a single site of action and is drug-resistant, with cyanoestrobin in a certain ratio can achieve a good synergistic effect in preventing staphylococci and potato blight and avoiding drug resistance. found. It has important application value in the production and application of pesticide varieties. So far, there have been no reports of a combination of the above two active ingredients.

Korean Patent No. 1945069

It is an object of the present invention to provide a bactericidal composition which has a good synergistic effect and can effectively delay the development of bactericide resistance.

The present invention achieves the above object through the following technical solutions:

The bactericidal composition, characterized in that the active ingredient of the bactericidal composition is oxathiapiprolin and cyanoestrobin, and the weight ratio is 5:1 to 1:1.

Preferably the weight ratio is 5:1 to 3:1;

More preferably, the weight ratio is 4:1.

In addition, the present invention further provides a bactericide, comprising 2 to 90% by weight of the bactericidal composition, and the remainder being an insecticide acceptable carrier and adjuvant.

Preferably the sterilizing agent comprises 6 to 70% by weight of said bactericidal composition;

More preferably, the sterilizing agent comprises 10 to 50% by weight of the bactericidal composition.

According to methods well known to those skilled in the art, the fungicide of the present invention may be in any agriculturally acceptable dosage form. Preferred dosage forms are emulsifiable concentrates, water dispersible granules, wettable powders, suspensions, microemulsions or water emulsions.

The present invention also provides a bactericidal emulsifiable concentrate, characterized in that the sterile emulsifiable concentrate comprises the following amounts of each component, calculated as weight percent:

Sterilization composition: 6-15%

Complex emulsifier: 15%

Co-solvent: 15%

remaining amount of water;

The complex emulsifier consists of Nongru 600# and Ningru 1600#, and the weight ratio is 2-5:1.

The cosolvent is one of ethanol, ethyl acetate, isopropanol, isobutanol and n-butanol, and the solvent is solvent oil No. 150.

Preferably, the weight ratio of the two complex emulsifiers is 4:1.

The bactericidal composition of the present invention may be provided in the form of a finished preparation, that is, in the form of a mixture of the substances of the composition; It may also be provided as a single dose formulation for direct mixing in the barrel (tank) prior to use. The concentrates of the present invention are generally mixed with water to obtain the desired active substance concentration.

The present invention also provides a sterilization method in which the bactericidal composition or fungicide of the present invention is applied to the crops and/or fruits and the places where they are grown or stored, either before or after the crops and/or fruits become ill.

The present invention also provides the use of said bactericidal composition or fungicide for the prevention and control of crop diseases. In particular, it is used to control grape downy mildew and potato blight.

Compared with the prior art, the present invention has the following advantageous effects:

(1) Compared with a single agent, the bactericidal composition of the present invention has a remarkable synergistic effect on disease, an improved bactericidal effect, and a co-toxicity coefficient of 135 or more.

(2) The composition is composed of active ingredients with different mechanisms of action, and the increase in the site of action is beneficial in overcoming and delaying the development of bacterial resistance.

(3) In addition, the bactericidal composition of the present invention can be prepared as an emulsifiable concentrate, so that the emulsification effect related to dispersibility, emulsification and stability characteristics can be greatly improved by selecting a specific complex emulsifier.

In order to make the object, technical solution, and advantage of the present invention clearer, the following specific examples are used for explanation, but the present invention is in no way limited to these examples. The following are only good examples for explaining the present invention and should not be construed as limiting the scope of the present invention. All modifications, equivalent substitutions or improvements made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.

I. Indoor Activity Examples

test drug

95% oxathiapiproline original drug.

95% cyanoestrobin.

1. Test method

1.1 Indoor Toxicity Test

1.1.1 Determination of Indoor Toxicity of Grape Downy Mildew

(1) Preparation of sporangia suspension

Collect raw leaves with grape downy mildew, wash the old mold layer of diseased leaves under running water using a brush, spread the leaves back onto a moisturizing tray with two layers of absorbent gauze at the bottom, and place the moisturizing tray with plastic film. sealed with Thereafter, it is placed in an artificial climate incubator under conditions of alternately culturing at 18° C., 90% relative humidity, and light and dark (light 12 hours, dark 12 hours). After new sporangia grew from the lesion, it was rinsed with distilled water and diluted to prepare a 1×10 ⁵ /L sporangia suspension for use.

(2) Indoor toxicity test

Drugs were administered 24 hours prior to inoculation. Another treatment solution was sprayed on the back side of a leaf disc with a diameter of 1.5 cm. After the liquid was air-dried, the back side of each treated leaf disc was placed on a Petri dish and the bottom of the Petri dish was covered with absorbent paper soaked in the same liquid. Fifteen leaf discs were placed concentrically on each dish and repeated 4 times for each dish. The blank control group was treated with 0.1% Tween-80 aqueous solution. Bacteria were inoculated after 24 h treatment. A pipette was used to inoculate 20 μL per leaf disc of the sporangial suspension in the center of the leaf disc. It was cultivated in an artificial climate incubator with a temperature of 22° C., a relative humidity of 90%, and the conditions of alternating light and dark (light 12 hours, dark 12 hours).

(3) Investigation method

Results were investigated when control was uniform. The severity of the disease was determined according to the ratio of the diseased spot area of the leaf disc to the area of the leaf disc, the number of diseased leaf discs and the severity of the disease were recorded, and the disease index and control effect were calculated according to the formula.

Disease severity grading criteria: Grade 0: no lesions; Grade 1: The area of the lesion accounts for less than 5% of the total leaf area; Grade 3: Lesions occupy 6%-25% of the total leaf area; Grade 5: The area of the lesion accounts for 26% to 50% of the total leaf area; Grade 7: The area of the lesion accounts for 51% to 75% of the total leaf area; Grade 9: The area of the lesion accounts for more than 76% of the total leaf area.

Disease index (%) = (∑ number of strains*representative value) / (total number of strains*highest representative value)*100

Control effect (%) = 1 - (Disease index of the treatment group after administration*Disease index of the control group before administration)/(Disease index of the treatment group before administration*Disease index of the control group after administration)*100

1.1.2 Determination of indoor toxicity of potato blight

A method of inhibiting mycelial growth was adopted: 0.5 mL of the drug solution and 4.5 mL of dissolved PDA medium containing 0.1% streptomycin sulfate were poured into a sterile Petri dish (diameter 5 cm) to make a medium plate containing the drug. After the medium was hardened, one potato-infecting bacteria cake (0.5 cm in diameter) was put on each side of the medium, and the side of the bacteria with mycelia was attached to the surface of the medium. Each treatment was repeated for 3 Petri dishes, and a sterile aqueous solution was used as a control. After incubation at 25° C. for 72 hours, the growth of the mycelium was recorded after treatment with compound A at different concentrations according to the growth of the mycelium in the control group. Using the cross method, the colony diameter (mm) was measured twice with a ruler and the average value was calculated. Mycelial growth inhibition rate was calculated according to the following formula.

Inhibition rate (%) = {(Net growth of mycelia in the control group - Net growth of mycelia in the treatment group)/Net growth of mycelia in the treatment group}*100

Note: net growth of mycelium = colony diameter - punch diameter

1.1.3 Data processing and analysis

The logarithm of drug concentration was taken as the independent variable x, and the probability of control effect or inhibition rate was taken as the dependent variable y for regression analysis to obtain the toxicity regression curve equation y=a+bx and the correlation coefficient r, and the oxa according to the regression equation EC ₅₀ and 95% confidence intervals for single doses of thiapiproline and cyanoestrobin were calculated.

1.2 Determination of Combined Toxicity of Fungicide Compounds

According to the results of the toxicity test of oxathiapiproline and cyanoestrobin single dose, the combination was formulated in 7:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2 volume ratios. got angry The antibacterial effect of the compound on potato blight and grape downy mildew was measured according to method 1.1, and the co-toxicity coefficient (CTC) was calculated by the Sun Yunpei (1960) method. CTC≥120 means synergistic effect; CTC≤80 means antagonistic effect; 80<CTC<120 means an additive effect.

Toxicity Index (TI) = (EC ₅₀ of standard formulation / EC ₅₀ of test formulation)*100%

Actual Toxicity Index (ATI) of the mixed formulation (M) = (EC ₅₀ of the standard formulation / EC ₅₀ of the mixed formulation)*100

Theoretical Toxicity Index (TTI) of Mixed Formulation M = (TI of Drug A*A% in Drug M) + (TI of Drug B*B% in Drug M)

Theoretical Toxicity Index (TTI) of Mixed Formulation M = (ATI/TTI)*100

1.3 Control effect

The control effects of grape downy mildew and potato late blight are shown in Tables 1 and 2.

Table 1: Grape Downy Mildew Control Effect

formulation name regression equation EC ₅₀ (mg/L) (95% confidence interval) correlation coefficient Co-toxicity coefficient Oxathiapiprolin y=1.7395x+8.1732 0.015 (0.0088-0.9962) 0.9962 Cyanoestrobin y=3.02x+1.2321 17.6870 (12.6194-24.7896) 0.9752 Oxathiapiprolin: Cyanoestrobin 5:1 y=2.9892x+10.92 0.0102 (0.0074-0.0148) 0.9847 175.34 Oxathiapiprolin: Cyanoestrobin 4:1 y=2.0594x+10.607 0.0019 (0.0012-0.0031) 0.9932 201.09 Oxathiapiprolin: Cyanoestrobin 3:1 y=2.0329x+8.8788 0.01204 (0.0077-0.01978) 0.9931 165.64 Oxathiapiprolin: Cyanoestrobin 2:1 y=2.3743x+9.3888 0.01427 (0.0094-0.2138) 0.9981 157.59 Oxathiapiprolin: Cyanoestrobin 1:1 y=1.3614x+7.261 0.02169 (0.0117-0.0408) 0.9933 138.27 Oxathiapiprolin: Cyanoestrobin 1:2 y=1.2539x+6.6984 0.0435 (0.0191-0.1025) 0.9914 100.35 Oxathiapiprolin: Cyanoestrobin 7:1 y=2.3118x+6.8575 0.1576 (0.1037-0.2385) 0.9894 107.99

As can be seen in Table 1, it can be seen that the synergistic effect is large when the weight ratio of oxathiapiproline:cyanoestrobin is 5:1 to 1:1, especially when the weight ratio is 4:1. When the weight ratio of the two is not in the range of 5:1 to 1:1, for example, when the weight ratio of the two is 7:1 or 1:2, the combination of the two shows only an additive effect.

Table 2: Potato blight control effect

formulation name regression equation EC ₅₀ (mg/L) (95% confidence interval) correlation coefficient Co-toxicity coefficient Oxathiapiprolin y=1.2825x+3.6389 11.5151 (5.9596-22.2493) 0.9936 Cyanoestrobin y=2.2192x+2.0337 24.7217 (16.0618-38.0674) 0.9926 Oxathiapiprolin: Cyanoestrobin 5:1 y=2.7601x+2.7618 6.4771 (4.5154-9.2709) 0.9933 195.23 Oxathiapiprolin: Cyanoestrobin 4:1 y=2.8351x+2.769 6.1228 (4.2851-8.7465) 0.9957 210.65 Oxathiapiprolin: Cyanoestrobin 3:1 y=2.0134x+3.1639 8.1127 (4.9763-13.3964) 0.9746 163.93 Oxathiapiprolin: Cyanoestrobin 2:1 y=1.3361x+4.9147 9.7619 (6.3061-4.9498) 0.9777 143.57 Oxathiapiprolin: Cyanoestrobin 1:1 y=2.5824x+2.246 11.6375 (7.9853-17.0056) 0.9928 135.06 Oxathiapiprolin: Cyanoestrobin 1:2 y=1.8848x+2.6081 18.3619 (11.5651-29.8517) 0.9985 97.42 Oxathiapiprolin: Cyanoestrobin 7:1 y=1.8626x+3.0309 11.3935 (6.7608-19.2458) 0.9995 108.34

As can be seen in Table 2, it can be seen that the synergistic effect is large when the weight ratio of oxathiapiproline:cyanoestrobin is 5:1 to 1:1, especially when the weight ratio is 4:1. When the weight ratio of the two was not in the range of 5:1 to 1:1, for example, when the weight ratio of the two was 7:1, the combination of the two showed only an additive effect. When the weight ratio of the two was 1:2, the combination of the two showed an antagonistic effect.

2. Preparation example

Preparation Example 1:

After adding 60 g of solvent oil No. 150 (process 180-210 ℃, effluent ≥ 98%, flash point ≥ 62 ℃) to a 200 ml flask, 15 g of isopropanol (analytical grade) was added and a glass rod for 5 minutes. stirred. To the mixed solution, 10 g of the sterilizing composition (weight ratio of oxathiapiproline to cyanoestrobin 2:1) was added and stirred with a glass rod for 10 minutes. 15 g of an emulsifier made by mixing Nongru 600# and Ningru 1600# (the weight ratio of the two is 2:1) was added to the mixed solution. After stirring for 5 minutes, a 10% oxathiapiproline and cyanoestrobin EC formulation was prepared.

Preparation Example 2:

It is the same as in Preparation Example 1 except that 15 g of an emulsifier made by mixing Nongru 600# and Ningru 1600# in a weight ratio of 5:1 was added.

Preparation Example 3:

It is the same as in Preparation Example 1 except that 15 g of an emulsifier made by mixing Nongru 600# and Ningru 1600# in a weight ratio of 4:1 was added.

Preparation Example 4:

It is the same as in Preparation Example 1 except that 15 g of an emulsifier made by mixing Nongru 600# and Ningru 1600# in a weight ratio of 1:1 was added.

Preparation Example 5:

The same as in Preparation Example 1 except that 15 g of an emulsifier made by mixing Nongru 600# and Ningru 1600# in a weight ratio of 6:1 was added.

Comparative Example 1:

The same as in Preparation Example 1 except that 15 g of Nongru 600# was added as an emulsifier.

Comparative Example 2:

The same as in Preparation Example 1 except that 15 g of Ningru 1600# was added as an emulsifier.

Comparative Example 3:

The same as in Preparation Example 1 except that 15 g of Nongru 500# was added as an emulsifier.

Comparative Example 4:

The same as in Preparation Example 1 except that 15 g of EL-40 was added as an emulsifier.

3. Example of emulsification effect test

1. The emulsion stability measurement method is in accordance with the GB/T1603-2001 method.

Standard hard water production (calcium carbonate standard hardness 342 mg L ^-1 )

0.304 g of anhydrous calcium chloride (accuracy 0.0002 g, hereinafter the same), 0.139 g of magnesium chloride, and crystalline water were placed in a 1000 ml beaker, dissolved in 600 ml of distilled water, and poured into a 1000 ml volumetric flask. Rinse the beaker with 300 ml distilled water, pour the rinse solution into a volumetric flask, fill up to the mark with distilled water, shake well and set aside.

100 ml of standard hard water at 25-30° C. was added to a 250 ml beaker, and 0.5 ml of emulsifiable concentrate samples (20-fold dilution) of Preparation Examples and Comparative Examples were obtained, and then slowly added to hard water with constant stirring. After adding the emulsifiable concentrate sample, stirring was continued for 30 seconds at a speed of 2-3 rps, immediately transfer the emulsion to a clean, dry 100 ml graduated cylinder and place the graduated cylinder in a (30±2) °C constant temperature water bath. was added for a period of time and the separation of the emulsion was observed. Emulsion stability is considered acceptable if there is no smooth oil (paste), oil settling, or sedimentation in the graduated cylinder.

2. Method for Determination of Emulsification and Dispersibility of Emulsion (Guo Wudi, 2003)

99.5 ㎖ of distilled water was put into a 100 ㎖ graduated cylinder, and 0.5 ㎖ of emulsifiable concentrates of Preparation Examples and Comparative Examples were transferred and the dispersion state was observed. After dropping the emulsifiable concentrate into the weighing container, the stopper was covered and the weighing container was rotated 15 times to observe the colostrum state. The evaluation criteria of emulsification and dispersibility are as follows.

Dispersibility criteria:

Excellent: Automatically dispersed in a blue fluorescent milky mist, which basically can automatically flip upwards without any noticeable particles, with a blue emulsion film on the wall.

Good: Most emulsifiable concentrates automatically disperse into a milky mist with visible particles or a small amount of smooth oil.

Yes: Can be dispersed as a milky white mist.

Poor: Not dispersed, oil droplets or particles settle.

Emulsification criteria:

Excellent: The emulsion is blue, transparent or translucent, with a strong milky white color.

Good: The emulsion is thick milky white or slightly bluish, with a milky white bottom and an emulsion film on the walls.

Yes: The emulsion is emulsified and has no gloss.

Poor: The emulsion is off-white with visible particles.

3. Test results

The test results are shown in Table 3 below.

Table 3 Emulsifying effect of various emulsifiers

emulsification effect dispersibility emulsifying stability Preparation Example 1 Excellent Good Qualified Preparation 2 Excellent Good Qualified Preparation 3 Excellent Excellent Qualified Preparation 4 Good Yes Qualified Preparation 5 Good Yes Qualified Comparative Example 1 Yes Yes Qualified Comparative Example 2 Good Yes unqualified Comparative Example 3 Good Yes unqualified Comparative Example 4 Good Poor unqualified

In the above table, it can be seen that when a complex emulsifier composed of Nongru 600# and Ningru 1600# is selected, that is, Preparation Examples 1-5 have better emulsification effect than Comparative Examples in which other emulsifiers or single agents are selected. The complex emulsifier itself is superior to other ratios when the weight ratio of the two is 2 to 5:1, and in particular, when the weight ratio of the two is 4:1, that is, the effect of Preparation Example 3 is the best. Oxathiapiproline and cyanoest The bactericidal composition of the present invention containing robin is illustrated by way of specific examples. Those skilled in the art can learn from the contents of the present invention, and can appropriately change the raw materials, process conditions and other links to achieve correspondingly different objects. Relevant modifications do not depart from the scope of the present invention. All similar substitutions and modifications will be apparent to those skilled in the art, and therefore all are considered to be included within the scope of the present invention.

Claims (5)

Use of a bactericidal composition for the prevention and control of potato blight, wherein the active ingredients of the bactericidal composition are oxathiapiprolin and cyanoestrobin in a weight ratio of 5:1 to 1:1.
The use according to claim 1, wherein the weight ratio is from 5:1 to 3:1.
The use according to claim 1, wherein the weight ratio is 4:1.
The use according to claim 1 , wherein the bactericidal composition is added together with pesticide acceptable carriers and adjuvants such that the weight of the bactericidal composition is from 2 to 90%.
5. Use according to claim 4, wherein the weight of the germicidal composition is between 6 and 70%.