阅读说明：本技术 一种含喹唑啉酮和春雷霉素的杀菌剂及其在防治植物病害中的应用 (Bactericide containing quinazolinone and kasugamycin and application of bactericide in preventing and treating plant diseases ) 是由 赵帅 韩奉娟 雍道敬 祝帅 尉莹莹 但丽霞 于 2020-12-09 设计创作，主要内容包括：本发明公开了一种含喹唑啉酮和春雷霉素的杀菌剂及其在防治植物病害中的应用。所述混配杀菌剂的活性成分由喹唑啉酮和春雷霉素组成,所述喹唑啉酮与所述春雷霉素的质量比为1～30：30～1。本发明混配杀菌剂可以制成悬浮剂、可溶性液剂、水乳剂或水分散粒剂等剂型。本发明混配杀菌剂能够防治植物病害,将所述混配杀菌剂进行喷雾或罐根,能够防治柑橘溃疡病、黄瓜细菌性角斑病、茄子枯萎病、茄子灰霉病等植物病害,稀释倍数优选低于1000倍。本发明混配杀菌剂的增效作用明显,提高了防治效果；两种杀菌剂的作用机理互异,扩大了组合物的防治谱；而且有利于减缓抗性的产生。(The invention discloses a bactericide containing quinazolinone and kasugamycin and application thereof in preventing and treating plant diseases. The active ingredients of the mixed bactericide comprise quinazolinone and kasugamycin, and the mass ratio of the quinazolinone to the kasugamycin is (1-30): 30 to 1. The mixed bactericide can be prepared into suspending agents, soluble liquid agents, aqueous emulsion or water dispersible granules and other dosage forms. The mixed bactericide can prevent and treat plant diseases, can prevent and treat the plant diseases such as citrus canker, cucumber bacterial angular leaf spot, eggplant wilt, eggplant gray mold and the like by spraying or canning the mixed bactericide, and the dilution multiple is preferably less than 1000 times. The synergistic effect of the mixed bactericide is obvious, and the control effect is improved; the action mechanisms of the two bactericides are different, so that the prevention and treatment spectrum of the composition is expanded; but also the resistance generation can be slowed down.)

1. A mixed bactericide comprises quinazolinone and kasugamycin as active ingredients;

the mass ratio of the quinazolinone to the kasugamycin is 1-30: 30 to 1.

2. The compounded germicide according to claim 1, wherein: the mass ratio of the quinazolinone to the kasugamycin is 1-12: 12 to 1.

3. The compounded germicide according to claim 1 or 2, characterized in that: the mixed bactericide is a suspending agent, a soluble liquid agent, an aqueous emulsion and water dispersible granules.

4. A compounded biocide as claimed in any one of claims 1 to 3, wherein: in the mixed bactericide, the sum of the mass percentages of the quinazolinone and the kasugamycin is 1-80%.

5. The compounded germicide according to claim 4, wherein: in the mixed bactericide, the sum of the mass percentages of the quinazolinone and the kasugamycin is 1-60%.

6. Use of a compound fungicide according to any one of claims 1-5 for controlling plant diseases.

7. Use according to claim 6, characterized in that: the plant diseases comprise citrus canker, cucumber bacterial angular leaf spot, eggplant wilt, eggplant gray mold and tomato gray mold.

8. Use according to claim 6 or 7, characterized in that: and spraying or canning the mixed bactericide.

Technical Field

The invention relates to a bactericide containing quinazolinone and kasugamycin and application thereof in preventing and treating plant diseases, belonging to the technical field of pesticides.

Background

The pesticide is an important agricultural production data, and the use of the pesticide is one of important means for ensuring high yield, high quality and high efficiency of agricultural production. At present, in agricultural pest control, the use of chemical pesticides accounts for more than 80%, but serious 3R (namely resistance, rampant and residue) problems are caused along with long-term use of the chemical pesticides. Therefore, the development of highly effective, environmentally friendly and low toxic chemical or biological pesticides is one of the important tasks in current research and development.

Quinazolinone belongs to quinoline alkaloids containing pyrimidine heterocycle, has stronger biological activity in the fields of medicine and pesticide, and mainly shows better effects of sterilization, disinsection and antivirus in the field of pesticide. The bactericide fluquinconazole and the acaricide fenazaquin with better effect on the market are developed from quinazolinone compounds. Kasugamycin is a metabolite produced by Streptomyces kasugaensis (Streptomyces kasugaensis), has strong systemic permeability and has prevention and treatment effects, and the action mechanism of the kasugamycin is to interfere the amino acid metabolism of a pathogenic bacterium esterase system, destroy the synthesis of protein, inhibit the growth of hypha and cause cell granulation, so that the pathogenic bacterium loses the reproductive and infection capacity, and the aims of killing the pathogenic bacterium and preventing and treating diseases are fulfilled. Therefore, it is necessary to compound quinazolinone and kasugamycin and study the control effect of the compound medicament.

Disclosure of Invention

The invention aims to provide a bactericidal agent containing quinazolinone and kasugamycin.

The active ingredients of the mixed bactericide provided by the invention consist of quinazolinone and kasugamycin;

the mass ratio of the quinazolinone to the kasugamycin is 1-30: 30-1;

preferably, the mass ratio of the quinazolinone to the kasugamycin is 1-12: 12-1 or 12-1: 1 or 12: 1.

the mixed bactericide can be prepared into suspending agents, soluble liquid agents, aqueous emulsion or water dispersible granules and other dosage forms.

In the mixed bactericide, the sum of the mass percentages of the quinazolinone and the kasugamycin can be 1-80%, preferably 1-60% or 1-26% or 26%, and the toxicity and the residue of the mixed bactericide reach better balance and the cost is lower.

The mixed bactericide can prevent and treat plant diseases, can prevent and treat the plant diseases such as citrus canker, cucumber bacterial angular leaf spot, eggplant wilt, eggplant gray mold, tomato gray mold and the like by spraying or canning the mixed bactericide, and the dilution multiple is preferably less than 1000 times.

The synergistic effect of the mixed bactericide is obvious, and the control effect is improved; the action mechanisms of the two bactericides are different, so that the prevention and treatment spectrum of the composition is expanded; but also the resistance generation can be slowed down.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 preparation of a suspension containing quinazolinone and kasugamycin

The weight percentage of each component is as follows: 2% of kasugamycin, 24% of quinazolinone, 3% of alkylphenol polyoxyethylene phosphate, 2% of polystyrene acrylate carboxylate copolymer, 1% of fatty alcohol polyoxyethylene ether, 0.2% of xanthan gum, 1% of magnesium aluminum silicate, 0.2% of sodium benzoate, 0.5% of polydimethylsiloxane emulsion and the balance of water which is used for supplementing to 100%.

The method comprises the following specific steps:

(1) adding water, an organic silicon defoaming agent, alkylphenol polyoxyethylene phosphate, a polystyrene acrylic carboxylate copolymer and fatty alcohol polyoxyethylene ether into a beaker, and uniformly dispersing;

(2) adding magnesium aluminum silicate, kasugamycin and quinazolinone into a beaker, and dispersing for 5min until the liquid medicine is homogeneous;

(3) transferring the liquid medicine to a sand mill for sand milling for 1.5 hours until the particle size is less than 5 mu m;

(4) adding xanthan gum and sodium benzoate into the liquid medicine, and uniformly stirring to obtain the suspension of the composition of kasugamycin and quinazolinone.

Example 2 preparation of a soluble solution containing quinazolinone and kasugamycin

The weight percentage of each component is as follows: 2% of kasugamycin, 24% of quinazolinone, 20% of xylene, 10% of phenethyl phenol polyoxyethylene ether, 5% of fatty alcohol polyoxyethylene ether, 0.2% of sodium benzoate and the balance of water for supplementing to 100%.

The method comprises the following specific steps:

(1) firstly, adding kasugamycin and quinazolinone into a beaker, then adding dimethylbenzene, starting stirring until the original drug is completely dissolved;

(2) adding the phenethyl phenol polyoxyethylene ether, the fatty alcohol polyoxyethylene ether and the sodium benzoate while stirring, continuously stirring until the phenethyl phenol polyoxyethylene ether, the fatty alcohol polyoxyethylene ether and the sodium benzoate are completely dissolved, and then adding water to obtain the soluble solution of the composition of the kasugamycin and the quinazolinone.

Example 3 preparation of an aqueous emulsion containing quinazolinone and kasugamycin

The weight percentage of each component is as follows: 2% of kasugamycin, 24% of quinazolinone, 3% of phenethyl phenol polyoxyethylene polyoxypropylene ether, 1% of fatty alcohol polyoxyethylene ether, 20% of xylene, 0.2% of sodium benzoate, 0.5% of an organic silicon defoamer and the balance of water to make up to 100%.

The method comprises the following specific steps:

(1) firstly, weighing kasugamycin and quinazolinone, adding the kasugamycin and the quinazolinone into a beaker, then adding dimethylbenzene, starting stirring, and stirring until the kasugamycin and the quinazolinone are completely dissolved to prepare an oil phase;

(2) adding phenethyl phenol polyoxyethylene polyoxypropylene ether, fatty alcohol polyoxyethylene ether, water, a preservative and a defoaming agent into a beaker, starting stirring, and stirring until the components are completely dissolved to prepare a water phase;

(3) adding the water phase into the oil phase, and stirring uniformly. And then shearing for 10min to obtain the composition aqueous emulsion of kasugamycin and quinazolinone.

Example 4 preparation of Water dispersible granules containing quinazolinone and kasugamycin

The weight percentage of each component is as follows: 2% of kasugamycin, 24% of quinazolinone, 2% of sodium dodecyl sulfate, 10% of alkyl naphthalene sulfonate, 5% of dispersing agent NNO and the balance of ammonium sulfate for supplementing to 100%.

The method comprises the following specific steps:

(1) uniformly mixing kasugamycin, quinazolinone, sodium lauryl sulfate, alkyl naphthalene sulfonate, a diffusant NNO and ammonium sulfate;

(2) crushing the uniformly mixed materials by airflow to 800 meshes;

(3) adding the crushed materials into a container, adding a small amount of water, and kneading;

(4) adding the kneaded material into an extrusion granulator, and carrying out extrusion granulation;

(5) putting the manufactured granules into a drying oven for drying;

(6) and sieving the dried particles, and screening out powder and the like which are not granulated to obtain the kasugamycin and quinazolinone composition water dispersible granules.

EXAMPLE 5 indoor bacteriostasis test of different formulations of quinazolinone and kasugamycin containing compositions against citrus canker

1. Reagent for testing

26% kasugamycin quinazolinone suspension, prepared according to example 1

26% kasugamycin quinazolinone solution, example 2 preparation

26% kasugamycin quinazolinone aqueous emulsion, example 3 preparation

26% kasugamycin quinazolinone water dispersible granule, example 4 preparation

2. Test method

A bacteriostatic test is carried out by adopting a filter paper method, 2mL of prepared bacterial suspension of citrus canker pathogen, namely citrus Xanthomonas citri subsp. Preparing the reagent to be tested into the required concentration, clamping a piece of sterile filter paper (diameter is 0.6cm) by using a sterile forceps, soaking the sterile filter paper into the liquid medicine, stopping the sterile filter paper for a moment at the edge of the container when the sterile filter paper is clamped out, filtering off the redundant liquid medicine, placing the filter paper in the center of a medicine-containing flat plate, slightly pressing the filter paper by using the forceps to ensure that the filter paper is fully and tightly attached to a culture medium, and measuring the diameter of a bacteriostatic ring after the culture dish is placed in a constant-temperature incubator at 28 ℃ for culturing for 48 hours. Each treatment was repeated 3 times, and a blank was made with a sterile filter paper sheet soaked in sterile water.

3. Test results

The test results are shown in table 1, when the content of kasugamycin in the composition is 2% and the content of quinazolinone in the composition is 24%, the difference of the antibacterial effect of the compositions of different dosage forms is large. When the test agent is diluted by 1000 times, the bacteriostatic effect of the soluble agent is optimal, and the effect of the soluble agent is not obviously different from that of an aqueous emulsion, but is obviously higher than that of a suspending agent and water dispersible granules. The diameter of the bacteriostatic circle of the suspending agent is 19.3mm, which is obviously higher than that of the water dispersible granules.

TABLE 126% kasugamycin-quinazolinone composition with different dosage forms has indoor bacteriostatic action on citrus canker pathogen

Note: letters after the same row of numbers indicate significant differences at P < 0.05 level (Duncan's New Bipolar Difference method).

EXAMPLE 6 indoor bacteriostasis test of suspensions containing quinazolinone and kasugamycin against Leptosphaeria citricola

1. Reagent for testing

2% kasugamycin solution from Qingdao Star brand crop science, Inc

24% quinazolinone suspension, prepared according to the method of example 1

26% kasugamycin quinazolinone suspension, prepared according to example 1

2. Test method

A bacteriostatic test is carried out by adopting a filter paper method, 2mL of the prepared suspension of the citrus canker pathogenic bacteria is added into a sterilized LB culture medium (45-60 ℃), the culture medium and the suspension are mixed uniformly by slight oscillation, and the culture dish is poured to prepare a bacteria-containing flat plate. Preparing the reagent to be tested into the required concentration, clamping a piece of sterile filter paper (diameter is 0.6cm) by using a sterile forceps, soaking the sterile filter paper into the liquid medicine, stopping the sterile filter paper for a moment at the edge of the container when the sterile filter paper is clamped out, filtering off the redundant liquid medicine, placing the filter paper in the center of a medicine-containing flat plate, slightly pressing the filter paper by using the forceps to ensure that the filter paper is fully and tightly attached to a culture medium, and measuring the diameter of a bacteriostatic ring after the culture dish is placed in a constant-temperature incubator at 28 ℃ for culturing for 48 hours. Each treatment was repeated 3 times, and a blank was made with a sterile filter paper sheet soaked in sterile water.

3. Test results

The test results are shown in table 2, when the test agent is diluted by 1000 × for treatment, the diameter of the bacteriostatic circle of the 24% kasugamycin-quinazolinone suspending agent is significantly larger than that of the bacteriostatic circle of the 2% kasugamycin aqueous solution and that of the 22% quinazolinone suspending agent, and the combination of the kasugamycin and the quinazolinone has a synergistic effect. When the 26% kasugamycin-quinazolinone suspending agent is diluted by 500 times, the diameter of the inhibition zone is 22.6mm, and has no significant difference with 1000 times of dilution, and when the 26% kasugamycin-quinazolinone suspending agent is diluted by 2000 times, the diameter of the inhibition zone has no significant difference with 22% quinazolinone suspending agent (1000 times).

Indoor bacteriostatic action of table 226% kasugamycin and quinazolinone suspending agent on citrus canker pathogen

Note: letters after the same row of numbers indicate significant differences at P < 0.05 level (Duncan's New Bipolar Difference method).

Example 7, 26% kasugamycin quinazolinone suspension indoor tests for inhibition of Botrytis cinerea

1. Reagent for testing

2% kasugamycin solution from Qingdao Star brand crop science, Inc

24% quinazolinone suspension, prepared according to the method of example 1

26% kasugamycin quinazolinone suspension, prepared according to example 1

2. Test method

Adopting a hypha growth rate method, preparing a reagent to be tested into a required concentration, uniformly mixing the reagent with a melted and cooled PDA culture medium (45-50 ℃), pouring a flat plate to prepare a medicine-containing flat plate, taking the PDA culture medium added with isometric sterile water as a control, and inoculating a Botrytis cinerea-Botrytis cinerea (Botrytis cinerea) cake with the diameter of 6mm in the center of the flat plate. When the control grows over 3/4 culture dishes, the cross method measures the colony diameters of the control group and the treatment group, calculates the bacteriostasis rate, and sets 3 times for each treatment.

3. Test results

The test results are shown in table 3.

TABLE 326 indoor bacteriostasis test of kasugamycin and quinazolinone suspending agent on Botrytis cinerea

Note: letters after the same row of numbers indicate significant differences at P < 0.05 level (Duncan's New Bipolar Difference method).

As can be seen from Table 3, the 26% kasugamycin-quinazolinone suspension treated at 500X showed the highest inhibition rate of 90.79% against Botrytis cinerea. The test agent is diluted by 1000 times, and the inhibition rate of the 26% kasugamycin-quinazolinone suspending agent on botrytis cinerea is obviously higher than that of a 2% kasugamycin aqueous solution and a 24% quinazolinone suspending agent. The control effect of the 26% kasugamycin and quinazolinone suspending agent of 2000 x on botrytis cinerea is 71.05%, and the treatment has no significant difference with the treatment of 2% kasugamycin aqueous solution of 1000 x and 24% quinazolinone suspending agent of 1000 x.

Example 8, 26% kasugamycin-quinazolinone suspension on Botrytis cinerea

1. Reagent for testing

2% kasugamycin solution from Qingdao Star brand crop science, Inc

24% quinazolinone suspension, prepared according to the method of example 1

26% kasugamycin quinazolinone suspension, prepared according to example 1

2. Test article and control object:

test work: eggplant of the species Bulita 702

The control object is: gray mold is caused by Botrytis cinerea (Botrytis cinerea)

3. Conditions of the test

The test site is arranged in an eggplant protection site in Shouguang city, Shandong province, and the test for controlling gray mold of eggplant is carried out in a greenhouse in the eggplant protection site in Shouguang city in 2020. The soil is sandy land. The disease is serious. The cultivation conditions and management measures of all the test cells are consistent.

4. Test design and arrangement

In the test, 6 treatments of 26% kasugamycin and quinazolinone suspending agent 500X, 1000X and 2000X, 2% kasugamycin soluble solution, 24% quinazolinone suspending agent 1000 times solution and no-application clear water are designed as controls, and each cell is 24m²The above steps are repeated for 4 times, and all the cells are arranged randomly.

5. Test investigation and calculation method

The disease condition base was investigated before the application of the drug, 7 days after the application of the drug, 15 days after the application of the drug, and 3 times of the whole test investigation. The investigation method adopts a random sampling method to carry out investigation. 5 plants are randomly selected in each cell, 2 branch number brands are selected for each plant, 5 leaves are counted from the branch tip to the lower part after the pro-drug is completely spread, and 50 leaves are investigated in each cell. Statistical analysis of the experimental data was performed using the Duncan new complex pole difference method. During the pesticide application period, the treatment is observed to have no phytotoxicity on the eggplants.

The grading method comprises the following steps:

level 0: no disease spots;

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 20 percent of the whole leaf area;

and 7, stage: the lesion area accounts for 21-50% of the whole leaf area;

and 9, stage: the lesion area accounts for more than 51% of the whole leaf area.

Calculating disease index and prevention and treatment effect according to the following formula:

6. results

6.1 prevention and treatment effects of test agent on Botrytis cinerea

Test results show that the control effects of the 26% kasugamycin-quinazolinone suspending agent diluted by 500 x and 1000 x times are respectively 91.60% and 91.30% 7 days after application, and are obviously higher than those of the other 3 treatments. When the 2% kasugamycin aqueous solution and the 24% quinazolinone suspending agent are diluted by 1000 times and the 26% kasugamycin-quinazolinone suspending agent is diluted by 2000 times, the control effect is equivalent, and no significant difference exists. 15 days after the drug is applied, the treatment control effect of 500 times dilution of 26% kasugamycin and quinazolinone suspending agent is the best, which is up to 91.07%, and the control effect of 1000 times dilution of 26% kasugamycin and quinazolinone suspending agent and 2000 times dilution of the 26% kasugamycin and quinazolinone suspending agent are not obviously different, but are obviously higher than the control effect of 1000 times dilution of 2% kasugamycin aqueous solution and 24% quinazolinone suspending agent.

TABLE 46 field efficacy test of kasugamycin-quinazolinone suspension on Botrytis cinerea

Note: letters after the same row of numbers indicate significant differences at P < 0.05 level (Duncan's New Bipolar Difference method).